\newtheorem{definition}{Definition}
-\title{Jalview 2.5: A manual and introductory tutorial }
+\title{Jalview 2.10.3: A manual and introductory tutorial }
\author{David Martin, James Procter, Andrew Waterhouse, Saif Shehata, Nancy Giang,Suzanne Duce and Geoff Barton}
\date{Manual version 1.2.3 6th May 2011}
{\Huge
-Jalview 2.9.0b2}
+Jalview 2.11}
\vspace{0.5in}
{\huge
Manual and Introductory Tutorial }
-\vspace{2.4in}
+\vspace{2.2in}
{\large
-David Martin, James Procter, Andrew Waterhouse, Saif Shehata, Nancy Giang,
+David Martin, James Procter, Ben Soares
+Andrew Waterhouse, Saif Shehata, Nancy Giang
-Suzanne Duce and Geoff Barton
-
+Mungo Carstairs, Charles Ofoegbu, Kira Mour\~{a}o
+
+Suzanne Duce and Geoff Barton
}
-\vspace{1.2in}
+\vspace{0.9in}
School of Life Sciences, University of Dundee
Dundee, Scotland DD1 5EH, UK
-
-
\vspace{2in}
-Manual Version 1.6
+Manual Version 1.9.3
% post CLS lifesci course on 15th January
% draft. Remaining items are AACon, RNA visualization/editing and Protein disorder analysis exercises.
-1st September 2016
+
+10th May 2019
\end{center}
\clearemptydoublepage
-% ($Revision$) 11th October 2010.}
+% ($Revision: 1.55 $) 11th October 2010.}
% TODO revise for 2.6
\pagenumbering{roman}
\section{Introduction}
\subsection{Jalview}
Jalview is a multiple sequence alignment viewer, editor and analysis tool.
-Jalview is designed to be platform independent (running on Mac, MS Windows, Linux
+It is designed to be platform independent (running on Mac, MS Windows, Linux
and any other platforms that support Java). Jalview is capable of editing and
analysing large alignments (thousands of sequences) with minimal degradation in
-performance, and able to show multiple integrated views of the alignment and
-other data. Jalview can read and write many common sequence formats including
+performance. It is able to show multiple integrated views of the alignment
+and other data. Jalview can read and write many common sequence formats including
FASTA, Clustal, MSF(GCG) and PIR.
There are two types of Jalview program. The {\bf Jalview Desktop} is a standalone
application that provides powerful editing, visualization, annotation and
-analysis capabilities. The {\bf JalviewLite} applet has the same core
+analysis capabilities. The {\bf JalviewJS} that has the same core
visualization, editing and analysis capabilities as the desktop, without the
-desktop's webservice and figure generation capabilities. It is designed to be
-embedded in a web page,\footnote{A demonstration version of Jalview (Jalview Micro
-Edition) also runs on a mobile phone but the functionality is limited to sequence
-colouring.} and includes a javascript API to allow customisable display of
-alignments for web sites such as Pfam.\footnote{\url{http://pfam.xfam.org}}
+desktop's webservice. It is designed to be
+opened in a web browser,\footnote{A demonstration version of Jalview (Jalview Micro
+Edition) also runs on a mobile phone but the functionality is limited.} and includes a javascript API.
-The Jalview Desktop in this version
-provides access to protein and nucleic acid sequence, alignment and structure
+The Jalview Desktop provides access to protein and nucleic acid sequence, alignment and structure
databases, and includes the Jmol\footnote{ Provided under the LGPL licence at
-\url{http://www.jmol.org}} viewer for molecular structures, and the VARNA\footnote{Provided under GPL licence at \url{http://varna.lri.fr}} program for the visualization of RNA secondary structure. A
-Distributed Annotation System (DAS) client\footnote{jDAS - released under Apache license (v2.0) at \url{http://code.google.com/p/jdas}} which facilitates the retrieval and display of third party sequence
-annotation in association with sequences and any associated structure. It also
-provides a graphical user interface for the multiple sequence alignment, conservation analysis and protein disorder prediction methods provided as {\bf Ja}va {\bf B}ioinformatics
-{\bf A}nalysis {\bf W}eb {\bf S}ervices (JABAWS). JABAWS\footnote{released under GPL at \url{http://www.compbio.dundee.ac.uk/jabaws}} is a system for running bioinformatics programs that you can download and run on your own machine or cluster, or install on compute clouds.
+\url{http://www.jmol.org}} and Chimera viewer for molecular structures, and the
+VARNA\footnote{Provided under GPL licence at \url{http://varna.lri.fr}} program for the visualization of RNA secondary structure. It also
+provides a graphical user interface for the multiple sequence alignment, conservation analysis
+and protein disorder prediction methods provided as {\bf Ja}va {\bf B}ioinformatics
+{\bf A}nalysis {\bf W}eb {\bf S}ervices (JABAWS). JABAWS\footnote{released under GPL at \url{http://www.compbio.dundee.ac.uk/jabaws}} is a system for
+running bioinformatics programs that you can download and run on your own machine or cluster, or install on compute clouds.
\subsection{Jalview's Capabilities}
% TODO add references to appropriate sections for each capability described here.
building, principal components analysis, physico-chemical property conservation
and sequence consensus analyses are built into the program. Web services enable
Jalview to access online alignment and secondary structure prediction programs,
-as well as to retrieve protein and nucleic acid sequences, alignments, protein structures and sequence annotation. Sequences, alignments, trees, structures, features and alignment annotation may also be exchanged with the local filesystem. Multiple visualizations of an alignment may be worked on simultaneously, and the user interface provides a comprehensive set of controls for colouring and layout. Alignment views are dynamically linked with Jmol structure displays, a tree viewer and spatial cluster display, facilitating interactive exploration of the alignment's structure. The application provides its own Jalview project file format in order to store the current state of an alignment and analysis windows. Jalview also provides WYSIWIG\footnote{WYSIWIG: What You See Is What You Get.} style figure generation capabilities for the preparation of alignments for publication.
+as well as to retrieve protein and nucleic acid sequences, alignments, protein structures and sequence annotation.
+Sequences, alignments, trees, structures, features and alignment annotation may also be exchanged with the local filesystem.
+Multiple visualizations of an alignment may be worked on simultaneously, and the user interface provides a comprehensive set of controls for colouring and layout.
+Alignment views are dynamically linked with Jmol and UCSF Chimera\footnote{UCSF Chimera needs to be installed separately. It is available free for academic use from \url{https://www.cgl.ucsf.edu/chimera/download.html}.} structure displays,
+a tree viewer and spatial cluster display, facilitating interactive exploration of the alignment's structure. The application provides its own Jalview project file format in order
+to store the current state of an alignment and analysis windows. Jalview also provides WYSIWIG\footnote{WYSIWIG: What You See Is What You Get.} style
+ figure generation capabilities for the preparation of alignments for publication.
\begin{figure}[htbp]
\begin{center}
\includegraphics[width=5.8in]{images/jvcapabilities.pdf}
\subsection{About this Tutorial }
This tutorial is written in a manual format with short exercises where
-appropriate, typically at the end of each section. This chapter concerns the
+appropriate. The first few sections concerns the
basic operation of Jalview and should be sufficient for those who want to
launch Jalview (Section \ref{startingjv}), open an alignment (Section
\ref{loadingseqs}), perform basic editing (Section
\ref{selectingandediting}), colouring (Section \ref{colours}), and produce
publication and presentation quality graphical output (Section \ref{layoutandoutput}).
-In addition, the manual covers the additional visualization and
-analysis techniques available in Jalview. This includes working
-with the embedded Jmol molecular structure viewer, building and viewing trees and PCA
-plots, and using trees for sequence conservation analysis. An overview of
-the Jalview Desktop's webservices is given in Section \ref{jvwebservices}, and
-the alignment and secondary structure prediction services are described
-in detail in Sections \ref{msaservices} and \ref{protsspredservices}. Section \ref{featannot} details the creation and visualization of sequence
-and alignment annotation, and the retrieval of sequences and annotation from
-databases and DAS Servers. Section \ref{workingwithnuc} discusses
-specific features of use when working with nucleic acid sequences, such as translation and linking to protein
-coding regions, and the display and analysis of RNA secondary structure.
+The remaining sections of the manual cover the visualization and
+analysis techniques available in Jalview. These include working
+with the embedded Jmol molecular structure viewer (or UCSF Chimera), building
+and viewing trees and Principal Components Analysis (PCA) plots, and using
+trees for sequence conservation analysis. Section \ref{featannot} details the creation and visualization of sequence,
+features and alignment annotation. The alignment and secondary structure prediction services are described
+in detail in Sections \ref{msaservices} and \ref{protsspredservices}
+respectively. Section \ref{workingwithnuc} discusses
+specific features of use when working with nucleic acid sequences, such as translation and linking to protein coding regions, and
+the display and analysis of RNA secondary structure. An overview of
+the Jalview Desktop's webservices is given in Section \ref{jvwebservices}.
%^Chapter \ref{jalviewadvanced} The third chapter covers the detail^ of Jalview and is aimed at the user who is
%already familiar with Jalview operation but wants to get more out of their
Keystrokes using the special non-symbol keys are represented in the tutorial by
enclosing the pressed keys with square brackets ({\em e.g.} [RETURN] or [CTRL]).
-Keystroke combinations are combined with a `-' symbol ({\em e.g.} [CTRL]-C means
-press [CTRL] and the `C' key) simultaneously.
+Keystroke combinations are denoted with a `-' symbol ({\em
+e.g.} [CTRL]-C means press [CTRL] and the `C' key simultaneously).
Menu options are given as a path from the menu
-that contains them - for example {\sl File $\Rightarrow$ Input Alignment
+that contains them. For example {\sl File $\Rightarrow$ Input Alignment
$\Rightarrow$ From URL} means to select the `From URL' option from the `Input
Alignment' submenu of a window's `File' dropdown menu.
This tutorial is based on the Jalview
Desktop application. Much of the information will also be useful for users of
-the JalviewLite applet, which has the same core editing, analysis and visualization capabilities (see the
-\href{http://www.jalview.org/examples/applets.html}{JalviewLite Applet Examples}
-page for examples). The Jalview Desktop, however, is much more powerful, and
+the JalviewJS, which has the same core editing, analysis and visualization capabilities. The Jalview Desktop, however, is much more powerful, and
includes additional support for interaction with external web services, and
production of publication quality graphics.
The Jalview Desktop can be run in two ways; as an application launched from the
web {\sl via} Java webstart, or as an application loaded onto your hard drive.
-The webstart version is launched from the pink `Launch Jalview Desktop
+The webstart version is launched from the `Launch Jalview Desktop
button' at the top right hand side of pages of the website
\href{http://www.jalview.org}{(www.jalview.org)}.
To download the locally installable version, follow the links on the download
%% this paragraph needs to be rewritten for the new signed certificate from Certum.
-When the application is launched with webstart, two dialogs may appear before
+When the application is launched with webstart, dialogs may appear before
the application starts. If your browser is not set up to handle webstart, then
clicking the launch link may download a file that needs to be opened
manually, or prompt you to select the program to handle the webstart
-file. If that is the case, then you will need to locate the {\bf javaws} program
+file. If that is the case, you will need to locate the {\bf javaws} program
on your system\footnote{The file that is downloaded will have a type of {\bf
application/x-java-jnlp-file} or {\bf .jnlp}. The {\bf javaws} program that can run
this file is usually found in the {\bf bin} directory of your Java
Jalview site. This behaviour can be switched off in the Jalview Desktop
preferences dialog by unchecking the open file option.
This alignment will look like the one in Figure \ref{startpage} (taken
-from Jalview version 2.7).
+from Jalview version 2.10.1).
%[figure 3 ]
\begin{figure}[htbp]
\exercise{Launching Jalview from the Jalview Website}{
\label{start}
-\exstep{Open the Jalview web
-site \href{http://www.jalview.org}{(www.jalview.org)}
+\exstep{Open the Jalview web site
+\href{http://www.jalview.org}{(www.jalview.org)}
in your web browser. Launch Jalview by clicking on the
pink `Launch Jalview' Desktop button in the top right hand corner. This
will download and open a jalview.jnlp webstart file.}
-\exstep {Dialogue boxes
+\exstep {Dialog boxes
will open and ask if you want to open the jalview.jnlp file as the file is an application downloaded from the
-Internet, click Open. (Note you maybe asked to update Java, if you agree then it
-will automatically update the Java software). As Jalview opens, four demo
+Internet, click {\sl Open}. (Note you may be asked to update Java, if you agree
+then it will automatically update the Java software). As Jalview opens, four demo
Jalview windows automatically load.}
\exstep {If
you are having trouble, it may help changing the browser you are using, as the browsers and
-it's version may affect this process.}
-\exstep{To deactivate the opening of the 4 demo sequences during the launch, go
+its version may affect this process.}
+\exstep{To disable opening of the demonstration project during the launch, go
to the {\sl Tools $\Rightarrow$
Preferences...} menu on the desktop. A `Preference'
-dialogue box opens, untick the box adjacent to the `Open file' entry in the
+dialog box opens, untick the box adjacent to the `Open file' entry in the
`Visual' preferences tab.
-Click OK to save the preferences.}
+Click {\sl OK} to save the preferences.}
\exstep{Launch another Jalview workbench from the web site by clicking on the
pink Launch button.
The example alignment should not be loaded as Jalview starts up.}
\exstep{To reload the original demo file select the
{\em File$\Rightarrow$ From URL} entry in the Desktop menu. Click on
-the URL history button on the right hand side of the dialog box to view the
-files, select exampleFile\_2\_7.jar, then click OK.}
-{\bf Note:} Should you want to reload the example alignment or load your own
-sequence during the launch process, then go
+the URL history button (a downward arrow on the right hand side of the dialog
+box) to view the files, select exampleFile\_2\_7.jar
+(\url{http://www.jalview.org/examples/exampleFile_2_7.jar})
+then click {\sl OK}.}
+\begin{list}{$\circ$}{\newline
+ \newline {\bf
+ Notes}}
+
+\item {To make Jalview display a different alignment when it is launched, then go
to the {\sl Tools $\Rightarrow$
-Preferences...} menu on the desktop. The tick the `Open file' entry of `Visual'
-preferences tab, type in the URL of the sequence you want to load. This
-file will load during the start up process.
-As the jalview.jnlp file launches Jalview on your desktop, you
-may want to move this from the downloads folder to another folder.
-Opening from this file will allow Jalview to be launched offline.
+Preferences...} menu on the desktop. Then tick the `Open file' entry of `Visual'
+preferences tab, type in the URL of the sequence you want to load.}
+\item {You may want to move the jalview.jnlp file from your {\bf downloads} to another folder.}
+\item {Opening Jalview via the jnlp file will also allow Jalview to be launched offline.}
+\end{list}
-{\bf Help launching Jalview is available in videos on the Getting Started page
-of the Jalview website at \url{http://www.jalview.org/Help/Getting-Started}.}}
+{\bf See the video at:
+\url{http://www.jalview.org/Help/Getting-Started}.}
+ }
\subsection{Getting Help}
\label{gettinghelp}
\subsubsection{Built in Documentation}
Jalview has comprehensive on-line help documentation. Select {\sl Help
-$\Rightarrow$ Documentation} from the main window menu and a new window will
-open (Figure \ref{help}). The appropriate topic can then be selected from the
+$\Rightarrow$ Documentation} from the main desktop window menu and a new window
+will open (Figure \ref{help}). The appropriate topic can then be selected from the
navigation panel on the left hand side. To search for a specific topic, click
the `search' tab and enter keywords in the box which appears.
\subsubsection{Email Lists}
-The Jalview Discussion list {\tt jalview-discuss@jalview.org} provides a forum
+The Jalview Discussion list ({\tt jalview-discuss@jalview.org}) provides a forum
for Jalview users and developers to raise problems and exchange ideas - any
problems, bugs, and requests for help should be raised here. The {\tt
jalview-announce@jalview.org} list can also be subscribed to if you wish to be
Archives and mailing list
subscription details can be found in the Jalview web site's \href{http://www.jalview.org/community}{community section}.
+
\section{Navigation}
\label{jvnavigation}
The major features of the Jalview Desktop are illustrated in Figure \ref{anatomy}. The alignment window is the primary window for editing and visualization, and can contain several independent views of the alignment being worked with. The other windows (Trees, Structures, PCA plots, etc) are linked to a specific alignment view. Each area of the alignment window has a separate context menu accessed by clicking the right mouse button.
Jalview has two navigation and editing modes: {\bf normal mode}, where
editing and navigation is performed using the mouse, and {\bf cursor mode}
- where editing and navigation are performed using the keyboard. The {\bf F2 key}
- is used to switch between these two modes. With a Mac as the F2 is
- often assigned to screen brightness, one may often need to type {\bf function
- [Fn] key with F2} function
- [Fn]-F2.
+ where editing and navigation are performed using the keyboard. The {\bf F2}
+ key is used to switch between these two modes.
+
+ {\em Note:} On MacBooks and other laptops with compact keyboards, you may need
+ to press the function key {\bf [Fn]} when pressing any of the numbered function
+ keys. So to toggle between keyboard and normal mode, press {\bf [Fn]-[F2]}.
+
\begin{figure}[htb]
\begin{center}
\includegraphics[width=6.5in]{images/jalview_anatomy.pdf}
-\caption{{\bf The anatomy of Jalview.} The major features of the Jalview Desktop Application are labeled.}
+\caption{{\bf The anatomy of Jalview.} The major features of the Jalview Desktop
+Application are labeled.}
% TODO: modify text labels to be clearer - black on grey and black border for clarity
\label{anatomy}
\end{center}
Each alignment view shown in the alignment window presents a window onto the
visible regions of the alignment. This means that with anything more than a few
residues or sequences, alignments can become difficult to visualize on the
- screen because only a small area can be shown at a time. It can help,
- especially when examining a large alignment, to have an overview of the whole
- alignment. Select {\sl View $\Rightarrow$ Overview Window} from the Alignment
- window menu bar (Figure \ref{overview}\footnote{the menu shown in this figure
- is from Jalview 2.2, later versions have more options.}).
+ screen because only a small area can be shown at a time. Here, it helps, to
+ have an overview of the whole alignment, especially when it is large.
+ Select {\sl View $\Rightarrow$ Overview Window} from the Alignment
+ window menu bar (Figure \ref{overview}).
% (Figure4)
\begin{figure}[htbp]
\begin{center}
\includegraphics[width=4.5in]{images/overview.pdf}
-\caption{{\bf Alignment Overview Window.} The overview window for a view is opened from the {\em View} menu.}
+\caption{{\bf Alignment Overview Window.} The overview window for a view is
+opened from the {\em View} menu.}
\label{overview}
\end{center}
\end{figure}
The red box in the overview window shows the current view in the alignment
window. A percent identity histogram is plotted below the alignment overview.
-Shaded parts indicate rows and columns of the alignment that are hidden (in this
-case, a single row at the bottom of the alignment - see Section
+Shaded parts indicate rows and columns of the alignment that are hidden (for more information see Section
\ref{hidingregions}). You can navigate around the alignment by dragging the red
box.
the alignments and analysis windows at once, then use the {\sl Window
$\Rightarrow$ Close All} option from the Jalview desktop.
-{\bf \em{Warning: make sure you have saved your work because this cannot be
+{\bf \em{Warning: Make sure you have saved your work because this cannot be
undone!}} }
\parbox[c]{3in}{\centerline{\includegraphics[width=2.5in]{images/start_closeall.pdf}
}}
Rapid movement to specific positions is accomplished as listed below:
\begin{list}{$\circ$}{}
\item {\bf Jump to Sequence {\sl n}:} Type a number {\sl n} then press [S] to
-move to sequence (row). {\sl n}
+move to sequence (row) {\sl n}.
\item {\bf Jump to Column {\sl n}:} Type a number {\sl n} then press [C] to move to column {\sl n} in the alignment.
\item {\bf Jump to Residue {\sl n}:} Type a number {\sl n} then press [P] to move to residue number {\sl n} in the current sequence.
\item {\bf Jump to column {\sl m} row {\sl n}:} Type the column number {\sl m}, a comma, the row number {\sl n} and press [RETURN].
%TODO insert a figure for the Find dialog box
\exercise{Navigation}{
+\label{navigationEx}
Jalview has two navigation and editing modes: {\bf normal} mode (where editing
and navigation are via the mouse) and the {\bf cursor} mode (where editing and
navigation are via the keyboard).
-The {\bf F2 key} is used to switch between these two modes. With a Mac often
-need to type function {\bf Fn key and F2}, as button is often assigned to
-screen brightness. Jalview always starts up in {\bf normal mode}.
+The {\bf F2} key is used to switch between these two modes. With a Mac, the key
+combination {\bf Fn and F2} keys are needed, as the {\bf F2} button is
+often assigned to screen brightness. Jalview always starts up in normal mode.
\exstep{Load an example alignment from its URL
(\url{http://www.jalview.org/examples/exampleFile_2_7.jar}) via the Desktop
-using {\sl File $\Rightarrow$ Input Alignment $\Rightarrow$ From URL} dialog
+using {\sl File $\Rightarrow$ Input Alignment $\Rightarrow$ from URL} dialog
box.
-(The URL should be stored in its history and clicking on the {\sl down arrow} is
-an easy way to access it.)}
+(The URL should be stored in its history and clicking on the {\sl down arrow}
+on the dialog box is an easy way to access it.)}
\exstep{Scroll around the alignment using the alignment (vertical) and ruler (horizontal) scroll bars.}
-\exstep{Find the Overview Window, {\sl Views
+\exstep{Find the Overview Window, {\sl View
$\Rightarrow$ Overview Window} and open it. Move around the
alignment by clicking and dragging the red box in the overview window.}
-\exstep{Return to the alignment window. Look at the status bar (lower left hand
-corner of the alignment window) as you move the mouse over the alignment. It indicates information about the
+\exstep{Return to the alignment window, look at the status bar (lower left
+hand corner of the alignment window) as you move the mouse over the alignment. It indicates information about the
sequence and residue under the cursor.}
-\exstep{Press [F2] key (or [Fn]/[F2] on Mac) to enter {\bf Cursor mode}. Use
+\exstep{Press [F2] key, or [Fn]-[F2] on Mac, to enter Cursor mode. Use
the direction keys to move the cursor around the alignment.}
\exstep{Move to sequence 7 by pressing {\bf 7 S}. Move to column 18 by pressing
{\bf 1 8 C}. Move to residue 18 by pressing {\bf 1 8 P}. Note that these can be
two different positions if gaps are inserted into the sequence. Move to sequence 5,
-column 13 by typing {\bf 1 3 , 5 [RETURN]}.}
+column 13 by typing {\bf 1 3 , 5} [RETURN].}
{\bf Note:} To view Jalview's comprehensive on-line help documentations select
-Help on desktop menu, clicking on Documentation will open a Documentation
-window. Select topic from the navigation panel on the left hand side or use the
-Search tab to select specific key words.
-{\bf Help navigating is available in videos on the Getting Started page
-of the Jalview website at \url{http://www.jalview.org/Help/Getting-Started}}
+{\sl Help} in desktop window menu, clicking on {\sl Documentation} will open a
+Documentation window. Select topics from the navigation panel on the left hand
+side or use the Search tab to locate specific key words.
+
+{\sl\bf See the video at:
+\url{http://www.jalview.org/Help/Getting-Started}.}
}
\section{Loading Sequences and Alignments}
%Jalview provides many ways to load your own sequences. %For this section of the
% tutorial you will need to download the file http://www.jalview.org/tutorial/alignment.fa to a suitable location on your hard drive.
\subsection{Drag and Drop}
- In most operating systems you can just drag a file icon from a file browser
- window and drop it on an open Jalview application window. The file will then be opened as a new alignment window. %You can try this with the tutorial file you have downloaded. When you have opened the file, close it again by selecting the close control on the window. If you drop an alignment file onto an open alignment window it will be appended.
+ In most operating systems you can drag a file icon from a file browser
+ window and drop it on an open Jalview application window. The file will then be opened as a new alignment window.
+ %You can try this with the tutorial file you have downloaded. When you have opened the file, close it again by selecting the close control on the window. If you drop an alignment file onto an open alignment window it will be appended.
Drag and drop also works when loading data from a URL -
-simply drag the link or url from the address panel of your browser on to an
+simply drag the link or url from the address panel of your browser onto an
alignment or the Jalview desktop background and Jalview will load data from the
URL directly.
% (Figure \ref{drag})
\subsection{From a File}
Jalview can read sequence alignments from a sequence alignment file. This is a
text file, {\bf not} a word processor document. For entering sequences from a
-wordprocessor document see Cut and Paste (Section \ref{cutpaste}) below. Select
+wordprocessor document see Cut and Paste (Section \ref{cutpaste}). Select
{\sl File $\Rightarrow$ Input Alignment $\Rightarrow$ From File} from the main
menu (Figure \ref{loadfile}). You will then get a file selection window where
you can choose the file to open. Remember to select the appropriate file type.
by Jalview. The way to read sequences from these documents is to select the
data from the document and copy it to the clipboard. There are two ways to
do this. One is to right-click on the desktop background, and select the
-`Paste to new alignment' option in the menu that appears. The other is to select
+`Paste to new window' option in the menu that appears. The other is to select
{\sl File $\Rightarrow$ Input Alignment $\Rightarrow$ From Textbox} from the
-main menu, and paste the sequences into the textbox window that will appear
-(Figure \ref{loadtext}). In both cases, presuming that they are in the right
-format, Jalview will happily read them into a new alignment window.
+main menu, paste the sequences into the text window that will appear, and select
+{\sl New Window} (Figure \ref{loadtext}). In both cases, presuming that they are
+in the right format, Jalview will happily read them into a new alignment window.
%[fig 8]
\begin{figure}[htbp]
\label{fetchseq}
Jalview can retrieve sequences and sequence alignments from the public databases
housed at the European Bioinformatics Institute, including Uniprot, Pfam, Rfam
-and the PDB, as well as any DAS sequence server registered at the configured DAS
-registry. Jalview's sequence fetching capabilities allow you to avoid having to
+and the PDB. Jalview's sequence fetching capabilities allow you to avoid having to
manually locate and save sequences from a web page before loading them into
Jalview. It also allows Jalview to gather additional metadata provided by the
source, such as annotation and database cross-references.
-Select {\sl File $\Rightarrow$ Fetch Sequence(s) \ldots} from the main menu and
-a window will appear (Figure \ref{loadseq}). Pressing the database selection
-button in the dialog box opens a new window showing all the database sources
-Jalview can access (grouped by the type of database). Once you've selected the
-appropriate database, hit OK close the database selection window, and then enter
-one or several database IDs or accession numbers separated by a semicolon and
-press OK. Jalview will then attempt to retrieve them from the chosen database.
-Example queries are provided for some databases to test that a source is
-operational, and can also be used as a guide for the type of accession numbers
-understood by the source.\footnote{Most DAS sources support {\em range queries}
-that can be used to download just a particular range from a sequence database
-record.}
+
+To begin retrieving data, select {\sl File $\Rightarrow$ Fetch Sequences \ldots}
+from the main menu. A window will then appear (Figure \ref{loadseq}) showing all
+the database sources Jalview can access (grouped by the type of database). Once
+you've selected the appropriate database by double clicking it or hitting OK, the
+database selection window will close and the sequence fetcher for that database
+will appear. You can then enter one or several database IDs or accession numbers
+separated by a semicolon and press OK. Jalview will then attempt to retrieve them
+from the chosen database. Example queries are provided for some databases to test
+that a source is operational, and can also be used as a guide for the type of
+accession numbers understood by the source.
% [fig 9]
\begin{figure}[htbp]
\begin{center}
\end{center}
\end{figure}
-\subsection{Memory Limits}
-\label{memorylimits}
-Jalview is a Java program. One unfortunate implication of this is that Jalview
-cannot dynamically request additional memory from the operating system. It is
-important, therefore, that you ensure that you have allocated enough memory to
-work with your data. On most occasions, Jalview will warn you when you have
-tried to load an alignment that is too big to fit in to memory (for instance,
-some of the PFAM alignments are {\bf very} large). You can find out how much
-memory is available to Jalview with the desktop window's {\sl $\Rightarrow$
-Tools $\Rightarrow$ Show Memory Usage} function, which enables the display of
-the currently available memory at the bottom left hand side of the Desktop
-window's background. Should you need to increase the amount of memory available
-to Jalview, full instructions are given in the built in documentation (opened by
-selecting {\sl Help $\Rightarrow$ Documentation}) and on the JVM memory
-parameters page (\url{http://www.jalview.org/jvmmemoryparams.html}).
\exercise{Loading Sequences}{
\label{load}
\exstep{Use {\sl Window $\Rightarrow$ Close All} from the Desktop window menu to
close all windows.}
-\exstep{{\bf Loading sequences from URL:} Selecting File {\sl $\Rightarrow$
-Input Alignment $\Rightarrow$ From URL} from the Desktop and enter \textsf{http://www.jalview.org/tutorial/alignment.fa} in the box.
+%% TODO: omit or combine this exercise
+%% NB. Edge (MS default browser) doesn't actually allow you to 'save' the page, apparently
+\exstep{{\bf Loading sequences from URL:} Selecting {\sl File $\Rightarrow$
+Input Alignment $\Rightarrow$ From URL} from the Desktop and enter \url{http://www.jalview.org/tutorial/alignment.fa} in the box.
+Click {\sl OK} to load the alignment.}
-Click OK to load the alignment.}
+%%TODO: omit or combine
\exstep{{\bf Loading sequences from a file:} Close all windows using the
-{\sl Window $\Rightarrow$ Close All} menu option from the Desktop.
+{\sl Window $\Rightarrow$ Close All} menu option from the Jalview desktop.
Then type the same URL (\url{http://www.jalview.org/tutorial/alignment.fa}) into
-your web browser and {\bf save} the file to your desktop.
+your web browser and save the fasta file (.fa) file to your desktop using {\sl File
+$\Rightarrow$ Save Page as}.
Open the file you have just saved in Jalview by selecting {\sl File
-$\Rightarrow$ Input Alignment $\Rightarrow$ From File} from the desktop menu and
-selecting this file.
-Click OK to load.}
+$\Rightarrow$ Input Alignment $\Rightarrow$ From File} from the desktop menu.
+Select the file and click {\sl OK} to load.}
+
\exstep{{\bf Loading sequences by `Drag and Drop' / `Cut and Paste':}
-(i) Select {\sl Desktop $\Rightarrow$ Window
-$\Rightarrow$ Close All}. Then drag the alignment.fa file from the desktop and
-drop it onto the Jalview window, the alignment should open.
+(i) Drag the alignment.fa file that you have just saved from its folder and
+drop it onto the Jalview desktop window, the alignment should open.
+
+(ii) Open
+\url{http://www.jalview.org/examples/estrogenReceptorProtein.fa} in a web
+browser. Test the differences
+between (a) dragging the URL directly from browser onto the Jalview
+desktop.
+(If the URL is downloaded, alternatively locate the file in your download
+directory and drag it onto the desktop.) (b) dragging the URL from
+browser onto the existing alignment.fa alignment window in the Jalview desktop.
+
+(iii) Open \url{http://www.jalview.org/examples/estrogenReceptorCdna.fa} in a web
+browser. Note that this is a cDNA file. Drag the URL from
+browser onto the estrogenReceptorProtein.fa protein alignment window in
+the Jalview desktop. A dialogue box opens asking `Would you like to open as split
+window, with cDNA and protein linked?' select `Split Window' option. A
+split window opens in the Jalview desktop.
+}
-Test the differences
-between (a) dragging the sequence onto the Jalview desktop and (b)
-dragging the sequence onto an existing alignment window.
+\exstep{{\bf The text editor:}
-(ii) Open \url{http://www.jalview.org/tutorial/alignment.fa} in a
-web browser. Drag the URL directly from browser onto Jalview desktop. (If
-the URL is downloaded, then locate the file in your
-download directory and open it in a text editor.)
+(i) Open the alignment.fa file using text editor.
+Copy the sequence text into the clipboard using [CTRL]-A and then [CTRL]-C.
-(iii) Open the alignment.fa file using text editor. Copy the sequence text from the
-file into the clipboard and paste it into the desktop
-background by right-clicking and selecting Paste to new alignment option.
+(ii) Move the mouse pointer onto the Jalview desktop window's background and right-click
+to open the context window. Select the {\sl Paste to New Window} menu option.
-(iv) In the text editor, copy the sequence text from
-alignment.fa into the clipboard (usually {\sl via} the browser's {\sl Edit
-$\Rightarrow$ Copy} menu option). In the Desktop menu, select {\sl File
+(iii) In the Jalview desktop menu, select {\sl File
$\Rightarrow$ Input Alignment $\Rightarrow$ From Textbox}.
-Paste the clipboard into the large window using the {\sl Edit $\Rightarrow$
-Paste} text box menu option. Click {\sl New Window} and the alignment will be
-loaded.}
-\exstep{{\bf Loading sequences from Public Database:} Select {\sl File
-$\Rightarrow$ Fetch Sequence(s)...} from the Desktop to open up new window
-called New Sequence Fetcher.
-Press database selection button (top of the dialog box), this opens
-another window called Select Database Retrieval Source showing all the database
-sources.
-
-Select the {\bf PFAM seed} database and click OK, then enter the accession
-number {\bf PF03460} and click OK. An alignment of about 174 sequences should
-load. These can be viewed using the Overview window accessible from {\sl View
-$\Rightarrow$ Overview Window.}
-Several database IDs can be loaded by using semicolons to separate them.}
-{\bf Help loading sequences is available in videos on the Getting Started page
-of the Jalview website at \url{http://www.jalview.org/Help/Getting-Started}.}
-}
+Paste the clipboard into the large window using [CTRL]-V. Click {\sl New Window}
+and the alignment will be loaded.}
+
+\exstep{{\bf Loading sequences from Public Database:} (i) Select {\sl File
+$\Rightarrow$ Fetch Sequences...} from the Jalview desktop menu. The {\sl
+Select Database Retrieval Source} dialog will open listing all the database
+sources. Select the {\bf PFAM seed} database and click {\sl OK}.
+
+%TODO fix ss appearing as RNA ss issue with secondary structure from pfam when downloading the Full alignment.
+
+(ii) The {\sl New
+Sequence Fetcher} window will open. Enter the accession number {\bf PF03460}
+and click {\sl OK}.
+An alignment of about 174 sequences should load.}
+\exstep{These can be viewed using the Overview window accessible from {\sl View
+$\Rightarrow$ Overview Window.}}
+{\bf See the video at:
+\url{http://www.jalview.org/Help/Getting-Started}} }
+
+\subsection{Memory Limits}
+\label{memorylimits}
+Jalview 2.11 and later will automatically maximise the amount of memory available,
+but if you are using an earlier version or launching Jalview in a specialised way
+you may need ensure that you have allocated enough memory to
+work with your data. On most occasions, Jalview will warn you when you have
+tried to load an alignment that is too big to fit in to memory (for instance,
+some of the PFAM alignments are {\bf very} large). You can find out how much
+memory is available to Jalview with the desktop window's {\sl $\Rightarrow$
+Tools $\Rightarrow$ Show Memory Usage} function, which enables the display of
+the currently available memory at the bottom left hand side of the Desktop
+window's background. Should you need to increase the amount of memory available
+to Jalview, full instructions are given in the built in documentation (opened by
+selecting {\sl Help $\Rightarrow$ Documentation}) and on the JVM memory
+parameters page (\url{http://www.jalview.org/jvmmemoryparams.html}).
\section{Saving Sequences and Alignments}
\label{savingalignments}
-\subsection{Saving Alignments} Jalview allows the
-current sequence alignments to be saved to file so they can be restored at a
-later date, passed to colleagues or analysed in other programs. From the
-alignment window menu select {\sl File $\Rightarrow$ Save As} and a dialog box
+\subsection{Saving Alignments} Jalview allows alignments to be saved to file
+in a variety of formats so they can be restored at a later date, passed to
+colleagues or analysed in other programs. From the alignment window menu select {\sl File $\Rightarrow$ Save As} and a dialog box
will appear (Figure \ref{savealign}). You can navigate to an appropriate
directory in which to save the alignment. Jalview will remember the last filename
and format used to save (or load) the alignment, enabling you to quickly save
-the file during or after editing by using the {\sl File $\Rightarrow$ Save} entry.
+the file during or after editing by using the {\sl File $\Rightarrow$ Save}
+entry. The {\sl File $\Rightarrow$ Output To Textbox} menu option allows the alignment to be copied and pasted into
+other documents or web servers.
-Jalview offers several different formats in which an alignment can be saved. The
-jalview format (.jar) is the only format which will preserve the colours,
-groupings and other additional information in the alignment. The other formats
+Jalview offers several different formats in which an alignment can be saved.
+Of these, only the jalview project format (.jar or .jvp) will preserve the colours, groupings and other additional information in the alignment. The other formats
produce text files containing just the sequences with no visualization information, although some
allow limited annotation and sequence features to be stored (e.g. AMSA).
-Unfortunately only Jalview program can read Jalview files. The {\sl File
-$\Rightarrow$ Output To Textbox} menu option allows the alignment to be copied and pasted into
-other documents or web servers.
+Unfortunately, as far as we are aware only Jalview can read Jalview
+project files.
%[fig 10]
\begin{figure}[htbp]
\begin{center}
-\parbox[c]{1.0in}{
-\includegraphics[width=1.0in]{images/saveas.pdf}
+\parbox[c]{3.0in}{
+\includegraphics[width=3in]{images/saveas.pdf}
}
-\parbox[c]{4in}{
-\includegraphics[width=4in]{images/saveas2.pdf}
+\parbox[c]{3in}{
+\includegraphics[width=3in]{images/saveas2.pdf}
}
\caption{{\bf Saving alignments in Jalview to disk.}}
\label{savealign}
just a single alignment (e.g. because you have calculated trees or multiple
different alignments) then save your work as a Jalview Project
file (.jvp).\footnote{Tip: Ensure that you have allocated plenty of memory to
-Jalview when working with large alignments in Jalview projects. See Section \protect
-\ref{memorylimits} above for how to do this.}
+Jalview when working with large alignments in Jalview projects. See Section
+\ref{memorylimits} for how to do this.}
From the main menu select {\sl File $\Rightarrow$ Save Project} and a file save
dialog box will appear. Loading a project will restore Jalview to exactly the
view at which the file was saved, complete with all alignments, trees,
annotation and displayed structures rendered appropriately.
-} \parbox[c]{2in}{ \centerline {
-\includegraphics[width=1.5in]{images/saveproj.pdf} }}
+} \parbox[c]{2.3in}{ \centerline {
+\includegraphics[width=2.2in]{images/saveproj.pdf} }}
\exercise{Saving Alignments}{
\label{save}
-\exstep{Launch Jalview, or use close all windows.
-Load the ferredoxin
-alignment from PFAM (seed) data base using the PFAM seed accession number
-PF03460 (see Exercise \ref{load}). } \exstep{
+\exstep{Launch Jalview afresh, or use {\sl Desktop $\Rightarrow$ Window
+$\Rightarrow$ Close all }.}
+\exstep{Load the ferredoxin
+alignment ({\bf PF03460}) from {\bf PFAM (seed)} (see Exercise
+\ref{load}).
+} \exstep{
Select {\sl File $\Rightarrow$ Save As} from the alignment window menu. Choose a
location into which to save the alignment and select your preferred format. All
formats except {\sl Jalview } jvp can be viewed in a normal text editor (e.g.
Notepad) or in a web browser.
-Enter a file name and click {\sl Save}.
-
-Check this file by closing all windows and opening it with Jalview, or by
+Enter a file name, select file type and click {\sl Save}.}
+\exstep{Check this file by closing all windows and opening it with Jalview, or by
browsing to it with your web browser.}
-\exstep{ Repeat the previous step saving the files in different file formats.}
+\exstep{Repeat the previous steps saving the files in different file formats.}
\exstep{Select {\sl File $\Rightarrow$ Output to Textbox $\Rightarrow$ FASTA}.
Select and copy this alignment to the clipboard using the textbox menu options
{\sl Edit $\Rightarrow$ Select All} followed by {\sl Edit $\Rightarrow$ Copy}.
}
\exstep{Ensure at least one alignment window is active in Jalview. Open the
overview window {\sl View $\Rightarrow$ Overview Window}
- and scroll red box to any part of the alignment.
+ and click and drag to move the red box to any part of the alignment.
Select {\sl File
-$\Rightarrow$ Save Project} from the main menu and save the project in a
-suitable folder.
+$\Rightarrow$ Save Project} from the desktop window menu and save the project in a
+suitable folder.}
-Close all windows and then load the project {\sl via} the {\sl File
-$\Rightarrow$ Load Project} menu option. Observe how all the windows and
-positions are exactly as they were when they were saved. }
-{\bf Help saving sequences is available in videos on the Getting Started page
-of the Jalview website at \url{http://www.jalview.org/Help/Getting-Started}.}}
+\exstep{Close all windows and then load the project {\sl via} the {\sl File
+$\Rightarrow$ Load Project} menu option. Observe how many of the windows
+reopen. Are they the same as when they were saved ? } {\bf See the video at:
+\url{http://www.jalview.org/Help/Getting-Started}.} }
\chapter{Selecting and Editing Sequences }
A selected region can be copied and pasted as a new alignment
using the {\sl Edit $\Rightarrow$ Copy} and {\sl Edit $\Rightarrow$ Paste $\Rightarrow$ To New
Alignment} in the alignment window menu options.
-{\bf To clear (unselect) the selection press the [ESC] (escape) key.}
+{\bf To clear (unselect) the selection press the [ESC] key.}
\subsection{Selecting Arbitrary Regions}
-To select part of an alignment, place the mouse at the top left corner of the region you wish to select. Press and hold the mouse button and drag the mouse to the bottom right corner of the chosen region then release the mouse button. A dashed red box appears around the selected region (Figure \ref{select}).
+To select part of an alignment, place the mouse at the top left corner of the region you wish to select. Press and hold the mouse button and drag the mouse to the bottom right corner of the chosen region then release the mouse button.
+A dashed red box appears around the selected region (Figure \ref{select}).
%[fig 12]
\begin{figure}[htbp]
This selects the entire column of the alignment. Ranges of positions from the
alignment ruler can also be selected by clicking on the first position and then
holding down the [SHIFT] key whilst clicking the other end of the selection.
-Discontinuous regions can be selected by holding down [CTRL] and clicking on positions to add to the column selection. Note that each [CTRL]-Click changes the current selected sequence region to that column, but adds to the column selection. Selected columns are indicated by red highlighting in the ruler bar (Figure \ref{selectcols}).
+Discontinuous regions can be selected by holding down [CTRL] and clicking on
+positions to add to the column selection. Note that each [CTRL]-Click (PC) or
+[CMD]-Click (Mac) changes the current selected sequence region to that column,
+it adds to the column selection.
+Selected columns are indicated by red highlighting in the ruler bar (Figure \ref{selectcols}).
%[fig 13]
\begin{figure}[htbp]
\end{center}
\end{figure}
+To select multiple complete sequences, click and drag the mouse down the
+sequence ID panel. The same techniques can be used as for columns above ([SHIFT]-Click for continuous and [CTRL]-Click (or {\sl
+[CMD]-Click} for Mac) to select discontinuous
+ranges of sequences (Figure \ref{selectrows}).
+%[fig 14]
+
\subsection{Selecting Sequences}
\begin{figure}[htb]
\begin{center}
\includegraphics[width=4in]{images/select3.pdf}
-\caption{{\bf Selecting multiple sequences in an alignment.} Use [CTRL] or [SHIFT] to select many sequences at once.}
+\caption{{\bf Selecting multiple sequences in an alignment.} Use [CTRL] or
+[SHIFT] to select many sequences at once.}
\label{selectrows}
\end{center}
\end{figure}
-To select multiple complete sequences, click and drag the mouse down the
-sequence ID panel. The same techniques as used for columns (above) can be used
-with [SHIFT]-Click for continuous and [CTRL]-Click to select discontinuous
-ranges of sequences (Figure \ref{selectrows}).
-%[fig 14]
+
\subsection{Making Selections in Cursor Mode}
$\Rightarrow$ Invert Sequence/Column Selection} in the alignment window.
Inverting the selection is useful when selecting large regions in an alignment,
simply select the region that is to be kept unselected, and then invert the selection.
-This may also be useful when hiding large regions in an alignment (see Section \ref{hidingregions}
-below).
+This may also be useful when hiding large regions in an alignment (see Section \ref{hidingregions}).
Instead of selecting the columns and rows that are to be hidden, simply select the
region that is to be kept
visible, invert the selection, then select {\sl View $\Rightarrow$ Hide
created which are labeled regions of the alignment. To create a group, first
select the region which is to comprise the group. Then click the right mouse
button on the selection to bring up a context menu. Select {\sl Selection
-$\Rightarrow$ Group $\Rightarrow$ Edit name and description of
+$\Rightarrow$ Edit New Group $\Rightarrow$ Edit name and description of
current group}\footnote{In earlier versions of Jalview, this entry was variously
`Group', `Edit Group Name', or `JGroupXXXXX' (Where XXXXX was some serial
-number).} then enter a name for the group in the dialogue box which appears.
+number).} then enter a name for the group in the dialog box which appears.
+
+By default the new group will have a box drawn around it. The appearance of the group can be changed (see Section \ref{colours}).
+This group will stay defined even when the selection is removed.
-By default the new group will have a box drawn around it. The appearance of the group can be changed (see Section \ref{colours} below). This group will stay defined even when the selection is removed.
+
+\section{Exporting the Current Selection}
+The current selection can be copied to the clipboard (in PFAM format). It can
+also be output to a textbox using the output functions in the pop-up menu
+obtained by right clicking the current selection. The textbox enables quick
+manual editing of the alignment prior to importing it into a new window (using
+the {\sl New Window} button) or saving to a file with the {\sl File
+$\Rightarrow$ Save as } pulldown menu option from the text box.
\exercise{Making Selections and Groups}{
\label{exselect}
-\exstep{Close windows.
-Load the ferredoxin alignment (PF03460 from PFAM seed database).
-Choose a residue and place the mouse
-cursor on it (Residue information will show in alignment window status
-bar)
-Click and drag the mouse to create a selection. As you drag, a red box
-will `rubber band' out to
+\exstep{Load the ferredoxin alignment ({\bf PF03460} from {\bf PFAM (seed)}).
+}
+\exstep{Selecting an arbitrary region. Choose a residue and place the mouse
+cursor on it (residue information will show in alignment window status
+bar).
+Click and drag the mouse to the bottom-right to create a selection. As you drag,
+a red box will `rubber band' out to
show the extent of the selection.
Release the mouse
button and a red box borders the selected region.
background and a red box appears around the selected sequence.
Hold down [SHIFT] and click another sequence ID a few positions above or below.
Note how the selection expands to include all the sequences between the two positions on which you clicked.
-Hold down [CTRL] and then click on several sequences ID's both selected and
+Hold down [CTRL] (or [CMD] on Mac) and then click on several sequences' IDs - both selected and
unselected. Note how unselected IDs are individually added to the selection and previously selected IDs are
individually deselected.}
-\exstep{ Select column by clicking on the Alignment Ruler. Note whole
-column is highlighted with a red box.
-Hold down [SHIFT] and click column beyond. Note the selection expands to include
-all the sequences between the two positions on which you clicked.}
-
-\exstep{ To selecting arbitrary regions in alignment, place the mouse at the top
-left corner of the region you wish to select. Press and hold the mouse button.
-Drag the mouse to the bottom right corner of the chosen region and release the
-mouse button and a dashed red box appears around the selected region}
-\exstep{Enter Cursor mode using [F2] (or [Fn]-F2 for Macs).
+\exstep{ Select columns by clicking on the Alignment Ruler. Note
+that the selected column is marked with a red box.
+Hold down [SHIFT] and click a column beyond. Note the selection expands to
+include all the sequences between the two positions on which you clicked.}
+
+\exstep{Enter Cursor mode using [F2], or [Fn]-F2 for Macs.
Navigate to column 59, row 1 by pressing {\bf 5 9 , 1 [RETURN]}.
Press {\bf Q} to mark this position.
Navigate to column 65, row 8 by pressing {\bf 6 5 , 8 [RETURN]}. Press {\bf M}
-to complete the selection. Note to clear the selection press the {\bf[ESC]}
+to complete the selection. Note to clear the selection press the [ESC]
key.}
-\exstep{To create a {\bf group} from the selected the region, click the
+\exstep{To create a group from the selected the region, click the
right mouse button when mouse is on the selection, this opens a
-pop-up menu in the alignment window.
+context menu in the alignment window.
Open the {\sl Selection $\Rightarrow$ Edit New Group $\Rightarrow$ Group Colour
-} menu and select `Percentage Identity'.
+} menu and select {\sl Percentage Identity}.
This will turn the selected region into a group and colour it accordingly.}
\exstep{Hold down [CTRL] and use the mouse to select and deselect sequences in
the alignment by clicking on their Sequence ID label. Note how the group expands
-to include newly selected sequences, and the `Percentage Identity' colouring changes. }
+to include newly selected sequences, and the Percentage Identity colouring changes. }
\exstep{ Another way to resize the group is by using the mouse to click and drag
the right-hand edge of the selected group.}
-\exstep{The current selection can be {\bf exported} and saved by right clicking
-on the text area to open the Sequence ID pop-up menu. Follow the menus and pick an
-output format (eg BLC) from the {\sl Selection $\Rightarrow$ Output to Textbox
-\ldots} submenu.
+\exstep{The current selection can be exported and saved, place mouse on the text
+area and right clicking the mouse to open the Sequence ID context menu.
+Select appropriate menu option and pick an output format (eg BLC) from the
+{\sl Selection $\Rightarrow$ Output to Textbox \ldots} submenu.
}
-\exstep{In the alignment output window, try manually editing the alignment,
-importing group into a new alignment window by clicking the [New Window] button to import the
-file into a new alignment window.}
-{\bf Additional help is available from videos on the Jalview website at \url{http://www.jalview.org/training/Training-Videos}.}
+\exstep{In the Alignment output window that opens, try manually editing the
+alignment before clicking the {\sl New Window} button. This opens the
+edited alignment in a new alignment window.} {\bf See the video at:
+\url{http://www.jalview.org/training/Training-Videos}.}
% more? change colouring style. set border colour.
}
-\section{Exporting the Current Selection}
-The current selection can be copied to the clipboard (in PFAM format). It can
-also be output to a textbox using the output functions in the pop-up menu
-obtained by right clicking the current selection. The textbox enables quick
-manual editing of the alignment prior to importing it into a new window (using
-the [New Window] button) or saving to a file with the {\sl File $\Rightarrow$
-Save As } pulldown menu option from the text box.
\section{Reordering an Alignment}
-Sequence reordering is simple. Highlight the sequences to be moved then press the up or down arrow keys as appropriate (Figure \ref{reorder}). If you wish to move a sequence up past several other sequences it is often quicker to select the group past which you want to move it and then move the group rather than the individual sequence.
+Sequence reordering is simple. Highlight the sequences to be moved then press the up or down arrow keys ($\uparrow$, $\downarrow$) as appropriate (Figure \ref{reorder}).
+If you wish to move a sequence up past several other sequences it is often quicker to select the group past which you want to move it and
+then move the group rather than the individual sequence.
\begin{figure}[htbp]
\begin{center}
\end{figure}
\exercise{Reordering the Alignment}{
-\exstep{Close all windows in Jalview from desktop. Load the ferredoxin alignment (e.g.the
-PFAM domain PF03460 from PFAM seed).
-Select one of the sequence in the sequence ID panel, use the up and down
+\label{reorderex}
+\exstep{Close windows.
+
+Load the ferredoxin alignment ({\bf PF03460} from {\bf PFAM (seed)}).
+}
+\exstep{Select one of the sequence in the sequence ID panel, use the up and down
arrow keys to alter the sequence's position in the alignment. (Note that
this will not work in cursor mode)}
\exstep{To select and move multiple
-sequences, use hold [SHIFT] and [CTRL], and select two sequences separated by
-one or more un-selected sequences. Note how multiple sequences are grouped
-together when they are re-ordered using the up and down arrow keys.}
-{\bf Additional help is available from videos on the Jalview website
-at \url{http://www.jalview.org/training/Training-Videos}.}}
+sequences, use hold [SHIFT], and select two sequences separated by
+one or more un-selected sequences, repeat using the [CTRL] key. Note how
+multiple sequences are grouped together when they are re-ordered using the up and down arrow keys.}
+{\bf See the video at: \url{http://www.jalview.org/training/Training-Videos}.}
+}
\section{Hiding Regions}
\label{hidingregions}
-It is sometimes convenient to exclude some sequences or residues in the alignment without actually deleting them. Jalview allows sequences or alignment columns within a view to be hidden, and this facility has been used to create the several different views in the example alignment file that is loaded when Jalview is first started (See Figure \ref{startpage}).
+It is sometimes convenient to exclude some sequences or residues in the alignment without actually deleting them.
+Jalview allows sequences or alignment columns within a view to be hidden, and this facility has been used to create
+the several different views in the example alignment file that is loaded when Jalview is first started (see Figure \ref{startpage}).
To hide a set of sequences, select them and right-click the mouse on the
selected sequence IDs to bring up the context pop-up menu. Select {\sl Hide
-Sequences} and the sequences will be concealed, with a small blue triangle indicating their position (Figure \ref{hideseq}). To unhide (reveal) the sequences, right click on the triangle and select {\sl Reveal Sequences} from the context menu.
+Sequences} and the sequences will be concealed, with a small blue triangle indicating their position (Figure \ref{hideseq}).
+To unhide (reveal) the sequences, right click on the triangle and select {\sl Reveal Sequences} from the context menu.
\begin{figure}[htbp]
\begin{center}
-\includegraphics[width=1.5in]{images/hide1.pdf}
-\includegraphics[width=2.5in]{images/hide2.pdf}
-\includegraphics[width=1.75in]{images/hide3.pdf}
-\caption{{\bf Hiding Sequences} Hidden sequences are represented by a small blue
-triangle in the sequence ID panel.}
+\includegraphics[width=1.9in]{images/hide1.pdf}
+\includegraphics[width=2.7in]{images/hide2.pdf}
+\includegraphics[width=1.8in]{images/hide3.pdf}
+\caption{{\bf Hiding Sequences} Hidden sequences are represented by a small
+blue triangle in the sequence ID panel.}
\label{hideseq}
\end{center}
\end{figure}
\begin{figure}[htbp]
\begin{center}
-\includegraphics[width=2in]{images/hide4.pdf}
+\includegraphics[width=1.7in]{images/hide4.pdf}
\includegraphics[width=3in]{images/hide5.pdf}
-\includegraphics[width=1in]{images/hide6.pdf}
+\includegraphics[width=1.2in]{images/hide6.pdf}
\caption{{\bf Hiding Columns} Hidden columns are represented by a small blue
triangle in the ruler bar.}
\label{hidecol}
It is often easier to select the region that you intend to work with, rather
than the regions that you want to hide. In this case, select the required region and use the {\sl View $\Rightarrow$ Hide
-$\Rightarrow$ All but Selected Region } menu entry, or press [Shift]+[Ctrl]+H
+$\Rightarrow$ All but Selected Region } menu entry, or press [SHIFT]-[CTRL]-H
to hide the unselected region.
\subsection{Representing a Group with a Single Sequence}
-Instead of hiding a group completely, it is sometimes useful to work with just one representative sequence. The {\sl $<$Sequence ID$>$ $\Rightarrow$ Represent group with $<$Sequence ID$>$ } option from the sequence ID pop-up menu enables this variant of the hidden groups function. The remaining representative sequence can be visualized and manipulated like any other. However, any alignment edits that affect the sequence will also affect the whole sequence group.
-
-\exercise{Hiding and Revealing Regions}{
-\exstep{Close all windows, open the PFAM accession PF03460. Select a
-contiguous set of sequences by clicking and dragging on the sequence ID panel.
-Right click on the selected sequence IDs to bring up the sequence ID pop-up
-menu, select {\sl Hide Sequences}.
-}
-\exstep{
-Right click on the blue triangle indicating hidden sequences and select {\sl
-Reveal Sequences} in the panel. (If you have hidden all sequences then you will
-need to use the alignment window menu option {\sl View $\Rightarrow$ Show $\Rightarrow$
-All Sequences.}) }
-\exstep{
-Repeat the process but use a non-contiguous set of sequences. Note that when
-multiple regions are hidden there are two options, {\sl Reveal Sequences} and {\sl Reveal All}.
-}
-\exstep{Repeat the above using columns to hide and reveal columns
-instead of sequences.}
-\exstep{Select a region of the alignment, then add in some additional columns to
-the selection, and experiment with the `Hide all but selected region' function
-in {\sl View $\Rightarrow$ Hide $\Rightarrow$ All but selected region.}}
-\exstep{Select some sequences and pick one to represent the rest by hovering
-the mouse over this sequence. Bring up the Sequence ID pop-up menu by right
-clicking and then select {\sl (Sequence ID name) $\Rightarrow$ Represent group
-with (Sequence ID name )}. To reveal these hidden sequences, right click on the
-Sequence ID and in the pop-up menu select Reveal All.}
-{\bf Additional help is available from videos on the Jalview
-website at \url{http://www.jalview.org/training/Training-Videos}}}
+Instead of hiding a group completely, it is sometimes useful to work with just one representative sequence.
+The {\sl $<$Sequence ID$>$ $\Rightarrow$ Represent group with $<$Sequence ID$>$ } option from the sequence ID pop-up menu enables this variant
+of the hidden groups function. The remaining representative sequence can be visualized and manipulated like any other.
+Note, any alignment edits that affect the sequence will also affect the whole
+sequence group.
\begin{figure}[htb]
\end{center}
\end{figure}
+%% TODO introduce select/hide by annotation here
+%% favor coverage of these core interactions (hide, show, select, reorder, multiple view)
+%% TODO - if columns are selected, then View->hide selected region hides *all* sequences, but it should leave the unselected columns visible !
+
+\exercise{Hiding and Revealing Regions}{
+\label{hidingex}
+\exstep{Load the ferredoxin alignment ({\bf PF03460} from {\bf PFAM (seed)}).
+}
+\exstep{Select a contiguous set of sequences by clicking and dragging on the sequence ID panel.
+Right click on the selected sequence IDs to bring up the sequence ID context
+menu, select {\sl Hide Sequences}.
+}
+\exstep{
+Right click on the blue triangle indicating hidden sequences and select {\sl
+Reveal Sequences} in the panel. (If you have hidden all sequences then you will
+need to use the alignment window menu option {\sl View $\Rightarrow$ Show $\Rightarrow$
+All Sequences.}) }
+\exstep{
+Repeat the process but use a non-contiguous set of sequences. Note that when
+multiple regions are hidden you can select either {\sl
+Reveal Sequences} to reveal the hidden sequences that were clicked, or {\sl
+Reveal All}.
+}
+\exstep{Repeat the above using columns to hide and reveal columns
+instead of sequences.}
+\exstep{Select a region of the alignment, and experiment with the {\sl Hide all
+but selected region} option in {\sl View $\Rightarrow$ Hide $\Rightarrow$ All
+but selected region.}} \exstep{Select some sequences, pick one to represent the rest by hovering
+the mouse over this sequence. Bring up the Sequence ID context menu by right
+clicking and select {\sl (Sequence ID name) $\Rightarrow$ Represent group
+with (Sequence ID name )}. To reveal these hidden sequences, right click on the
+Sequence ID and in the context menu select {\sl Reveal All}.}
+{\bf See the video at: \url{http://www.jalview.org/training/Training-Videos}.}
+}
+
+
+
\section{Introducing and Removing Gaps}
-The alignment view provides an interactive editing interface, allowing gaps to be inserted or deleted to the left of any position in a sequence or sequence group. Alignment editing can only be performed whilst in keyboard editing mode (entered by pressing [F2]) or by clicking and dragging residues with the mouse when [SHIFT] or [CTRL] is held down (which differs from earlier versions of Jalview).
+The alignment view provides an interactive editing interface, allowing gaps to be inserted or deleted to the left of any position
+in a sequence or sequence group. Alignment editing can only be performed whilst in keyboard editing mode (entered by pressing [F2])
+ or by clicking and dragging residues with the mouse when [SHIFT], [CTRL] or [CMD] (Mac) is held down (which differs from earlier versions of Jalview).
+
+
+\subsection{Undoing Edits}
+Alignment edits can be undone {\sl via} the {\sl Edit $\Rightarrow$ Undo Edit}
+alignment window menu option, or [CTRL]-Z. An edit, if undone, may be re-applied
+with {\sl Edit $\Rightarrow$ Redo Edit}, or [CTRL]-Y. Note, however, that the
+{\sl Undo} function only works for edits to the alignment or sequence ordering.
+Colouring of the alignment, showing and hiding of sequences or modification of
+annotation that only affect the alignment's display cannot
+be undone.
\subsection{Locked Editing}
\label{lockededits}
\subsection{Introducing Gaps in a Single Sequence}
To introduce a gap, first select the sequence in the sequence ID panel and
then place the cursor on the residue to the immediate right of where the gap
-should appear. Hold down the SHIFT key and the left mouse button, then drag the sequence to the right until the required number of gaps has been inserted.
+should appear. Hold down the [SHIFT] key and the left mouse button, then drag the sequence to the right until the required number of gaps
+has been inserted.
-One common error is to forget to hold down [SHIFT]. This results in a selection which is one sequence high and one residue long. Gaps cannot be inserted in such a selection. The selection can be cleared and editing enabled by pressing the [ESC] key.
+One common error is to forget to hold down [SHIFT]. This results in a selection which is one sequence high and one residue long.
+Gaps cannot be inserted in such a selection. The selection can be cleared and editing enabled by pressing the [ESC] key.
\subsection{Introducing Gaps in all Sequences of a Group}
To insert gaps in all sequences in a selection or group, select the
required sequences in the sequence ID panel and then place the mouse cursor on
-any residue in the selection or group to the immediate right of the position in which a gap should appear. Hold down the CTRL key and the left mouse button, then drag the sequences to the right until the required number of gaps has appeared.
+any residue in the selection or group to the immediate right of the position in which a gap should appear. Hold down the [CTRL] key
+and the left mouse button, then drag the sequences to the right until the required number of gaps has appeared.
Gaps can be removed by dragging the residue to the immediate right of the gap
leftwards whilst holding down [SHIFT] (for single sequences) or [CTRL] (for a group of sequences).
% % better idea to introduce hiding sequences, and use the invert selection, hide
% others, to simplify manual alignment construction
+
+
+\subsection{Editing in Cursor Mode}
+Gaps can be easily inserted when in cursor mode (toggled with [F2]) by
+pressing [SPACE]. Gaps will be inserted at the cursor, shifting the residue
+under the cursor to the right. To insert {\sl n} gaps type {\sl n} and then
+press [SPACE]. To insert gaps into all sequences of a group, use [CTRL]-[SPACE]
+or [SHIFT]-[SPACE] (both keys held down together).
+
+Gaps can be removed in cursor mode by pressing [BACKSPACE]. First make sure you
+have everything unselected by pressing [ESC]. The gap under the cursor will be
+removed. To remove {\sl n} gaps, type {\sl n} and then press [BACKSPACE]. Gaps
+will be deleted up to the number specified. To delete gaps from all sequences of
+a group, press [CTRL]-[BACKSPACE] or [SHIFT]-[BACKSPACE] (both keys held down
+together). Note that the deletion will only occur if the gaps are in the same
+columns in all sequences in the selected group, and those columns are to the
+right of the selected residue.
+
+
\exercise{Editing Alignments}
%\label{mousealedit}
% TODO: VERIFY FOR 2.6.1 and 2.7 - NUMBERING/INSTRUCTIONS APPEAR OFF
-{You are going to manually reconstruct part of the example Jalview
+{
+\label{editingalignex}
+You are going to manually reconstruct part of the example Jalview
alignment available at
- \href{http://www.jalview.org/examples/exampleFile.jar}
- {http://www.jalview.org/examples/exampleFile.jar}.
-Remember to use [CTRL]+Z to undo an edit, or the {\sl File $\Rightarrow$
+ \href{http://www.jalview.org/examples/exampleFile.jvp}
+ {http://www.jalview.org/examples/exampleFile.jvp}.
+
+{\sl {\bf Mac Users:} Please use the Apple or [CMD] key in place of [CTRL]
+for key combinations such as [CTRL]-A. }
+
+Remember to use [CTRL]-Z to undo an edit, or the {\sl File $\Rightarrow$
Reload } function to revert the alignment back to the original version if you
want to start again.
-If you are using OSX, and a key combination - such as [CTRL]+A - does
- not work, then try pressing the [CMD] key instead of [CTRL].
\exstep{ Load the URL
\textsf{http://www.jalview.org/tutorial/unaligned.fa} which contains part of the
ferredoxin alignment from PF03460.}
-\exstep{ Select the first 7 sequences, and press H to hide them (or right click
-on the sequence IDs to open the sequence ID pop-up menu, and select {\sl Hide
+\exstep{ Select the first 7 sequences, and press H key to hide them (or right click
+on the sequence IDs to open the sequence ID context menu, and select {\sl Hide
Sequences}).}
\exstep{ Select FER3\_RAPSA and FER\_BRANA. Slide the sequences to
-the right so the initial {\bf A} lies at column 57 using the $\Rightarrow$ key.}
+the right so the initial residue A lies at column 57 using the $\rightarrow$
+key.}
\exstep{ Select FER1\_SPIOL, FER1\_ARATH, FER2\_ARATH, Q93Z60\_ARATH and
O80429\_MAIZE
-(Hint: you can do this by pressing [CTRL]-I to invert the sequence selection and then
-deselect FER1\_MAIZE), and use the $\Rightarrow$ key to slide them to so they
-begin at column 5 of the alignment view.}
+
+{\sl Hint: you can do this by pressing [CTRL]-I to invert the sequence selection and then
+deselect FER1\_MAIZE), and use the $\rightarrow$ key to slide them to so they
+begin at column 5 of the alignment view.}}
\exstep{ Select all the visible
sequences (those not hidden) in the block by pressing [CTRL]-A.
Insert a single
gap in all selected sequences at column 38 of the alignment by holding [CTRL]
-and clicking on the R at column 38 in the FER1\_SPIOL, then drag one
+and clicking on the residue R at column 38 in the FER1\_SPIOL, then drag one
column to right.
Insert another gap at column 47 in all sequences in the same way.}
\exstep{ Correct the ferredoxin domain alignment for FER1\_SPIOL by
-inserting two additional gaps after the gap at column 47: First press [ESC] to
+inserting two additional gaps after the gap at column 47. First press [ESC] to
clear the selection, then hold [SHIFT] and click and drag on the G and move it
two columns to the right.}
\exstep{ Now complete the
-alignment of FER1\_SPIOL with a {\bf locked edit} by pressing [ESC] and select
+alignment of FER1\_SPIOL with a locked edit by pressing [ESC] and select
columns 47 to 57 of the FER1\_SPIOL row. Move the mouse onto the G at column 50,
hold [SHIFT] and drag the G in column 47 of FER1\_SPIOL to the left by one
column to insert a gap at column 57.}
column 44, and insert 4 gaps at column 47 (after AAPM).}
\exstep{ Hold [SHIFT] and drag the I at column 39 of FER1\_MAIZE 2 columns to the
-right. Remove the gap at FER1\_MAIZE column 49 by [SHIFT]+click and drag left by
+right. Remove the gap at FER1\_MAIZE column 49 by [SHIFT]-click and drag left by
one column. Press [ESC] to clear the selection, and then insert three gaps in
FER1\_MAIZE at column 47 by holding [SHIFT] and click and drag the S in FER1\_MAIZE to the right by three columns. Finally,
remove the gap in O80429\_MAIZE at column 56 using [SHIFT]-drag to the left on
\exstep{ Use the
{\sl Edit $\Rightarrow$ Undo Edit} and {\sl Edit $\Rightarrow$ Redo Edit} menu
-option, or their keyboard shortcuts ([CTRL]+Z and [CTRL]+Y) to step
+option, or their keyboard shortcuts ([CTRL]-Z and [CTRL]-Y) to step
backwards and replay the edits you have made.}
}
-\subsection{Editing in Cursor mode}
-Gaps can be easily inserted when in cursor mode (toggled with [F2]) by
-pressing [SPACE]. Gaps will be inserted at the cursor, shifting the residue
-under the cursor to the right. To insert {\sl n} gaps type {\sl n} and then
-press [SPACE]. To insert gaps into all sequences of a group, use [CTRL]-[SPACE]
-or [SHIFT]-[SPACE] (both keys held down together).
-
-Gaps can be removed in cursor mode by pressing [BACKSPACE]. First make sure you
-have everything unselected by pressing ESC. The gap under the cursor will be
-removed. To remove {\sl n} gaps, type {\sl n} and then press [BACKSPACE]. Gaps
-will be deleted up to the number specified. To delete gaps from all sequences of
-a group, press [CTRL]-[BACKSPACE] or [SHIFT]-[BACKSPACE] (both keys held down
-together). Note that the deletion will only occur if the gaps are in the same
-columns in all sequences in the selected group, and those columns are to the
-right of the selected residue.
-
-\section{Undoing Edits}
-Jalview supports the undoing of edits {\sl via} the {\sl Edit $\Rightarrow$ Undo Edit}
-alignment window menu option, or CTRL-Z. An edit, if undone, may be re-applied
-with {\sl Edit $\Rightarrow$ Redo Edit}, or CTRL-Y. Note, however, that the
-{\sl Undo} function only works for edits to the alignment or sequence ordering.
-Colouring of the alignment, showing and hiding of sequences or modification of
-annotation cannot be undone.
\exercise{Keyboard Edits}
-{This continues on from exercise
-\ref{mousealedit}, and recreates the final part of the example ferredoxin
+{
+\label{keyboardsex}
+This continues on from the previous exercise, and recreates the final part of the example ferredoxin
alignment from the unaligned sequences using Jalview's keyboard editing mode.
-{\bf Note:} For Mac users, [CTRL]-[SPACE] command
-has the same effect as the [SHIFT]-[SPACE] command mentioned in this exercise.
-Window users should use [SHIFT]-[SPACE] rather than the [CTRL]-[SPACE] command,
-as this command will close the window.
+{\bf Window users:} Please {\em only use} [SHIFT]-[SPACE] in this
+exercise.
+
+{\bf Mac users:} [CTRL]-[SPACE] can also be used instead of [SHIFT]-[SPACE].
\exstep{Load the sequence alignment at
\textsf{http://www.jalview.org/tutorial/unaligned.fa}, or continue using the
-edited alignment from exercise \ref{mousealedit}. If you continue from the
+edited alignment. If you continue from the
previous exercise, first right click on the sequence ID panel and select
-{\sl Reveal All}. Enter cursor mode by pressing [F2].}
+{\sl Reveal All}. Enter cursor mode by pressing [F2] (or [Fn]-[F2] (Mac)).}
% TODO: BACKSPACE or DELETE WHEN SEQS ARE SELECTED WILL DELETE ALL SEQS JAL-783
\exstep{Insert 58 gaps at the start of the sequence 1 (FER\_CAPAA). Press
column 38.
Press {\sl 34C [BACKSPACE] 38C 2 [SPACE]}. Delete three gaps at column 44 and
insert one at column 47 by pressing {\sl 44C 3 [BACKSPACE] 47C [SPACE]}. The top five sequences are
-now aligned.}
-}
+now aligned.}}
+
\chapter{Colouring Sequences and Figure Generation}
\label{colouringfigures}
Colouring sequences is a key aspect of alignment presentation. Jalview allows
you to colour the whole alignment, or just specific groups. Alignment and
group colours are rendered
-{\sl below} any other colours, such as those arising from sequence features
+{\bf below} any other colours, such as those arising from sequence features
(these are described in Section \ref{featannot}). This means that if you
try to apply one of the colourschemes described in this section, and nothing
appears to happen, it may be that you have sequence feature annotation
displayed, and you may have to disable it using the {\sl View $\Rightarrow$
-Show Features} option before you can see your colourscheme.
+Show Sequence Features} option before you can see your colourscheme.
There are two main types of colouring styles: {\bf simple static residue}
colourschemes and {\bf dynamic schemes} which use conservation and consensus
analysis to control colouring. {\bf Hybrid colouring} is also possible, where
-static residue schemes are modified using a dynamic scheme. The individual schemes are described in Section \ref{colscheme} below.
+static residue schemes are modified using a dynamic scheme. The individual schemes are described in Section \ref{colscheme}.
\subsection{Colouring the Whole Alignment}
Selections or groups can be coloured in two ways. The first is {\sl via} the Alignment Window's {\sl Colour} menu as stated above,
after first ensuring that the {\sl Apply Colour To All Groups} flag is {\bf
not} selected.
- This must be turned {\sl off} specifically as it is {\sl on} by default.
+ This must be turned {\bf off} specifically as it is {\bf on} by default.
When unticked, selections from the Colours menu will only change the colour for residues in the current selection,
or the alignment view's ``background colourscheme'' when no selection exists.
\includegraphics[width=2in]{images/colourcons1.pdf}
\includegraphics[width=2in]{images/colourcons3.pdf}
\includegraphics[width=2in]{images/colourcons5.pdf}
-\caption{{\bf Conservation Shading} The density of the ClustalX style residue colouring is controlled by the conservation threshold. The effect of 0\% (left), 50\% (center) and 100\% (right) thresholds are shown.
+\caption{{\bf Conservation Shading} The density of the ClustalX style residue
+colouring is controlled by the conservation threshold. The effect of 0\% (left), 50\% (center) and 100\% (right) thresholds are shown.
}
\label{colcons}
\end{center}
\label{colscheme}
Full details on each colour scheme can be found in the Jalview on-line help. A brief description of each one is provided below:
-\subsubsection{ClustalX}
+\subsubsection{Clustalx}
\parbox[c]{3.5in}{This is an emulation of the default colourscheme used for alignments in ClustalX, a graphical interface for the ClustalW multiple sequence alignment program. Each residue in the alignment is assigned a colour if the amino acid profile of the alignment at that position meets some minimum criteria specific for the residue type. }
\parbox[c]{3in}{\includegraphics[width=2.75in]{images/col_clustalx.pdf}}
-\subsubsection{Blosum62}
+\subsubsection{Blosum62 Score}
\parbox[c]{3.5in}{Gaps are coloured white. If a residue matches the consensus sequence residue at that position it is coloured dark blue. If it does not match the consensus residue but the Blosum62 matrix gives a positive score, it is coloured light blue.}
\parbox[c]{3in}{
\subsubsection{Percentage Identity}
\parbox[c]{3.5in}{
-The Percent Identity option colours the residues (boxes and/or text) according to the percentage of the residues in each column that agree with the consensus sequence. Only the residues that agree with the consensus residue for each column are coloured.
+The Percentage Identity option colours the residues (boxes and/or text) according to the percentage of the residues in each column that agree with the consensus sequence. Only the residues that agree with the consensus residue for each column are coloured.
}
\parbox[c]{3in}{
\includegraphics[width=2.75in]{images/col_percent.pdf}
\subsubsection{Zappo}
\parbox[c]{3.5in}{
-The residues are coloured according to their physicochemical properties. The physicochemical groupings are Aliphatic/hydrophobic, Aromatic, Positive, Negative, Hydrophillic, conformationally special, and Cyst(e)ine.
+The residues are coloured according to their physicochemical properties. The physicochemical groupings are Aliphatic/hydrophobic, Aromatic,
+Positive, Negative, Hydrophillic, Conformationally special, and Cyst(e)ine.
}
\parbox[c]{3in}{
\includegraphics[width=2.75in]{images/col_zappo.pdf}
sequences and alignments.
} \parbox[c]{3in}{ \includegraphics[width=2.75in]{images/col_nuc.pdf} }
-\subsubsection{Purine Pyrimidine}
+\subsubsection{Purine/Pyrimidine}
\parbox[c]{3.5in}{ Residues are coloured according to whether the corresponding
nucleotide bases are purine (magenta) or pyrimidine (cyan) based. All non ACTG
residues are uncoloured. For further information about working with nucleic acid
} \parbox[c]{3in}{ \includegraphics[width=2.75in]{images/col_purpyr.pdf} }
-\subsubsection{RNA Helix Colouring}
+\subsubsection{By RNA Helices}
\parbox[c]{3.5in}{ Columns are coloured according to their assigned RNA helix as
defined by a secondary structure annotation line on the alignment. Colours for
each helix are randomly assigned, and option only available when an RNA
} \parbox[c]{3in}{
\includegraphics[width=2.75in]{images/col_rnahelix.pdf} }
-\exercise{Colouring Alignments}{
-\exstep{Ensuring that the {\sl Apply Colour To All Groups} flag is not selected
-in View sequence alignment menu.
- This must be turned {\sl off} specifically as it is {\sl on} by default.}
-\exstep{
-Open a sequence alignment, for example the PFAM domain PF03460 in PFAM seed.
-Select the alignment menu option {\sl Colour $\Rightarrow$ ClustalX}. Note the colour change. Now try all the other colour schemes in the {\sl Colour} menu. Note that some colour schemes do not colour all residues.
-}
-\exstep{
-Colour the alignment using {\sl Colour $\Rightarrow$ Blosum62}. Select a group
-of around 4 similar sequences. Use the context menu (right click on the group)
-option {\sl Selection $\Rightarrow$ Edit New Group $\Rightarrow$ Group Colour
-$\Rightarrow$ Blosum62} to colour the selection. Notice how some residues which
-were not coloured are now coloured. The calculations performed for dynamic
-colouring schemes like Blosum62 are based on the group being coloured, not the
-whole alignment (this also explains the colouring changes observed in exercise
-\ref{exselect} during the group selection step).
-}
-\exstep{
-Keeping the same selection as before, colour the complete alignment except
-the group using {\sl Colour $\Rightarrow$ Taylor}.
-Select the menu option {\sl Colour $\Rightarrow$ By Conservation}.
-Slide the selector from side to side and observe the changes in the alignment colouring in the selection and in the complete alignment.
-}
-}
-
\subsubsection{User Defined}
-This dialogue allows the user to create any number of named colour schemes at will. Any residue may be assigned any colour. The colour scheme can then be named. If you save the colour scheme, this name will appear on the Colour menu (Figure \ref{usercol}).
+This dialog allows the user to create any number of named colour schemes at
+will. Any residue may be assigned any colour. The colour scheme can then be
+named. If you save the colour scheme, this name will appear on the Colour menu
+(Figure \ref{usercol}).
\begin{figure}[htbp]
\begin{center}
-\includegraphics[width=2.5in]{images/col_user1.pdf}
+\includegraphics[width=2.1in]{images/col_user1.pdf}
\includegraphics[width=2in]{images/col_user2.pdf}
-\includegraphics[width=1.75in]{images/col_user3.pdf}
-\caption{{\bf Creation of a user defined colour scheme.} Residue types are assigned colours (left). The profile is saved (center) and can then be accessed {\sl via} the {\sl Colour} menu (right).}
+\includegraphics[width=2.1in]{images/col_user3.pdf}
+\caption{{\bf Creation of a user defined colour scheme.} Residue types are
+assigned colours (left). The profile is saved (center) and can then be accessed {\sl via} the {\sl Colour} menu (right).}
\label{usercol}
\end{center}
\end{figure}
+\exercise{Colouring Alignments}{
+\label{color}
+Note: Ensure that the {\sl Apply Colour
+To All Groups} flag is not selected in {\sl Colour} menu in the alignment window.
+% patch needed for 2.10 This must be turned {\sl off} specifically as it is on
+% by default.
+
+\exstep{Open a sequence alignment, for example the PFAM domain PF03460 in PFAM
+seed database. Select the alignment menu option {\sl Colour $\Rightarrow$
+Clustalx} and note the colour change. Now try all the other colour schemes in the {\sl Colour} menu.
+Note that some colour schemes do not colour all residues.}
+\exstep{Colour the alignment using {\sl Colour $\Rightarrow$ Blosum62 score}. Select a group
+of around 4 similar sequences. Use the context menu (right click on the group)
+option {\sl Selection $\Rightarrow$ Edit New Group $\Rightarrow$ Group Colour
+$\Rightarrow$ Blosum62 score} to colour the selection. Notice how some residues which
+were not coloured are now coloured. The calculations performed for dynamic
+colouring schemes like Blosum62 are based on the selected group,
+not the whole alignment. (This also explains the colouring changes observed in exercise
+\ref{exselect} during the group selection step).}
+\exstep{Keeping the same selection as before, colour the complete alignment except
+the group using {\sl Colour $\Rightarrow$ Taylor}.
+Select the menu option {\sl Colour $\Rightarrow$ By Conservation}.
+Slide the selector in the Conservation Colour Increment dialog box from side
+to side and observe the changes in the alignment colouring in the selection and in the complete alignment.}
+Note: Feature colours overlay residue colouring. The features colours can be
+toggled off by going to {\sl View $\Rightarrow$ Show Sequence Features}.
-\exercise{User Defined Colour Schemes}{
-\exstep{Load a sequence alignment. Select the alignment menu option {\sl Colour $\Rightarrow$ User Defined}. A dialogue window will open.
-}
-\exstep{Click on an amino acid button, then select a colour for that amino acid. Repeat till all amino acids are coloured to your liking.
-}
-\exstep{ Insert a name for the colourscheme in the appropriate field and click {\sl Save Scheme}. You will be prompted for a file name in which to save the colour scheme. The dialogue window can now be closed.
-}
-\exstep{
-The new colour scheme appears in the list of colour schemes in the {\sl Colour} menu and can be selected in future Jalview sessions.
-}
+{\bf See the video at: \url{http://www.jalview.org/training/Training-Videos}.}
}
+
+\exercise{User Defined Colour Schemes}{
+\label{colouex}
+\exstep{Load a sequence alignment PF03460 from the PFAM
+seed database. Ensure that the {\sl Colour $\Rightarrow$
+None} is selected. Select the alignment menu option {\sl Colour $\Rightarrow$
+User Defined}.
+A dialog window will open.}
+\exstep{Click on an amino acid button, then select a colour for that amino acid. Repeat till all amino acids are coloured to your liking.}
+\exstep{ Insert a name for the colourscheme in the appropriate field and click {\sl Save Scheme}. You will be prompted for a file name in which to save the colour scheme. The dialog window can now be closed.}
+\exstep{The new colour scheme appears in the list of colour schemes in the {\sl Colour} menu and can be selected in future Jalview sessions.
+
+{\bf See the video at:
+\url{http://www.jalview.org/training/Training-Videos}.}} }
+
+\exercise{Alignment Layout}{
+\label{exscreen}
+\exstep{Start Jalview and open the URL \textsf{http://www.jalview.org/examples/exampleFile.jvp}.
+Select {\sl Format $\Rightarrow$ Wrap} from the alignment window menu.
+Experiment with the various options from the {\sl Format} menu, for example adjust the ruler placement,
+sequence ID format and so on. }
+\exstep{Hide all the annotation rows by toggling {\sl Annotations $\Rightarrow$
+Show Annotations} from the alignment window menu. Reveal the annotations by selecting the same menu option.} \exstep{Deselect {\sl Format $\Rightarrow$ Wrap}. Right click on the
+annotation row labels to bring up the context menu, then select {\sl
+Hide This Row}. Bring up the context menu again and select {\sl
+Show All Hidden Rows} to reveal them.}
+\exstep{Annotations can be reordered by clicking on the annotations name and
+dragging the row to the desired position. Click on the {\sl Consensus} row and drag it upwards to just above {\sl Quality}.
+The rows should now be reordered. Features and annotations are covered in more detail in Section \ref{featannot}.}
+\exstep{Move the mouse to the top left hand corner of the annotation labels -
+a up/down arrow symbol should appear - when this is shown, the height of the {\sl Annotation Area} can be changed
+by clicking and dragging this icon up or down.}
+\bf See the video at:
+\url{http://www.jalview.org/training/Training-Videos}.}
+
\section{Formatting and Graphics Output}
\label{layoutandoutput}
Jalview is a WYSIWIG alignment editor. This means that for most kinds of graphics output,
\subsection{Multiple Alignment Views}
-Jalview is able to create multiple independent visualizations of the same underlying alignment - these are called {\sl Views}. Because each view displays the same underlying data, any edits performed in one view will update the alignment or annotation visible in all views.
+Jalview is able to create multiple independent visualizations of the same underlying alignment - these are called {\bf Views}.
+Because each view displays the same underlying data, any edits performed in one view will update the alignment or annotation visible in all views.
-\parbox[c]{4in}{Alignment views are created using the {\sl View $\Rightarrow$ New View} option of the alignment window or by Pressing [CTRL]-T. This will create a new view with the same groups, alignment layout and display options as the current one. Pressing G will gather together Views as named tabs on the alignment window, and pressing X will expand gathered Views so they can be viewed simultaneously in their own separate windows. To delete a group, press [CTRL]-W.}\parbox[c]{2.75in}{
+\parbox[c]{4in}{Alignment views are created using the {\sl View $\Rightarrow$
+New View} option of the alignment window or by pressing [CTRL]-T.
+This will create a new view with the same groups, alignment layout and display options as the current one.
+Pressing G key will gather together Views as named tabs on the alignment window, and pressing X key will expand gathered Views so they can be viewed
+simultaneously in their own separate windows.} \parbox[c]{2.75in}{
\begin{center}\centerline{
\includegraphics[width=2.5in]{images/mulv_tabs.pdf}}
\end{center}
}
+To delete a view, press [CTRL]-W (or [CMD]-W (Mac)).
+To rename a view, right click the view's name, this open the Enter View Name dialogue box, enter the desired name.
% JBPNote make an excercise on views ?
\subsection{Alignment Layout}
-Jalview provides two screen layout modes, unwrapped (the default) where the alignment is in one long line across the window, and wrapped, where the alignment is on multiple lines, each the width of the window. Most layout options are controlled by the Format menu option in the alignment window, and control the overall look of the alignment in the view (rather than just a selected region).
+Jalview provides two screen layout modes, unwrapped (the default) where the alignment is in one long line across the window, and wrapped,
+where the alignment is on multiple lines, each the width of the window. Most layout options are controlled by the Format menu option in the
+alignment window, and control the overall look of the alignment in the view (rather than just a selected region).
\subsubsection{Wrapped Alignments}
-Wrapped alignments can be toggled on and off using the {\sl Format $\Rightarrow$ Wrap} menu option (Figure \ref{wrap}). Note that the annotation lines are also wrapped. Wrapped alignments are great for publications and presentations but are of limited use when working with large numbers of sequences.
+Wrapped alignments can be toggled on and off using the {\sl Format $\Rightarrow$ Wrap} menu option (Figure \ref{wrap}). Note that
+the annotation tracks are also wrapped. Wrapped alignments are great for publications and presentations but are of limited use when
+working with large numbers of sequences.
-If annotations are not all visible in wrapped mode, expand the alignment window to view them. Note that alignment annotation (see Section \ref{featannot}) cannot be interactively created or edited in wrapped mode, and selection of large regions is difficult.
+If annotations are not all visible in wrapped mode, expand the alignment window to view them. Note that alignment annotation
+(see Section \ref{featannot}) cannot be interactively created or edited in wrapped mode, and selection of large regions is difficult.
\begin{figure}[htbp]
\begin{center}
\parbox[c]{2in}{\includegraphics[width=2in]{images/wrap1.pdf}}
\subsubsection{Fonts}
-\parbox[c]{4in}{The text appearance in a view can be modified {\sl via} the {\sl Format $\Rightarrow$ Font\ldots} alignment window menu. This setting applies for all alignment and annotation text except for that displayed in tool-tips. Additionally, font size and spacing can be adjusted rapidly by clicking the middle mouse button and dragging across the alignment window.}
+\parbox[c]{4in}{The text appearance in a view can be modified {\sl via} the {\sl Format $\Rightarrow$ Font\ldots} alignment window menu. This setting
+applies for all alignment and annotation text except for that displayed in tool-tips. Additionally, font size and spacing can be adjusted rapidly by
+clicking the middle mouse button and dragging across the alignment window.}
\parbox[c]{2in}{\centerline{\includegraphics[width=1.75in]{images/font.pdf}}}
\subsubsection{Numbering and Label Justification}
-Options in the {\sl Format} menu are provided to control the alignment view, and provide a range of options to control the display of sequence and alignment numbering, the justification of sequence IDs and annotation row column labels on the annotation rows shown below the alignment.
+Options in the {\sl Format} menu are provided to control the alignment view, and provide a range of options to control the display of sequence and
+alignment numbering, the justification of sequence IDs and annotation row column labels on the annotation rows shown below the alignment.
\subsubsection{Alignment and Group Colouring and Appearance}
-The display of hidden row/column markers and gap characters can be turned off with {\sl Format $\Rightarrow$ Hidden Markers} and {\sl Format $\Rightarrow$ Show Gaps}, respectively. The {\sl Text} and {\sl Colour Text} option controls the display of sequence text and the application of alignment and group colouring to it. {\sl Boxes } controls the display of the background area behind each residue that is coloured by the applied coloursheme.
+The display of hidden row/column markers and gap characters can be turned off with {\sl Format $\Rightarrow$ Show Hidden Markers} and
+{\sl Format $\Rightarrow$ Show Gaps}, respectively. The {\sl Text} and {\sl Colour Text} option controls the display of sequence text and the application of alignment and group colouring to it. {\sl Boxes } controls the display of the background area behind each residue that is coloured by the applied coloursheme.
\subsubsection{Highlighting Nonconserved Symbols}
The alignment layout and group sub-menu both contain an option to hide
display gaps or sequence symbols that differ from the consensus for each
column, and render all others with a `.'.
%TODO add a graphic to illustrate this.
+
\subsection{Annotation Ordering and Display}
% TODO: describe consensus, conservation, quality user preferences, and group
% annotation preferences.
The annotation lines which appear below the sequence alignment are described in
-detail in Section \ref{featannot}. They can be hidden by toggling the {\sl View
-$\Rightarrow$ Show Annotations} menu option. Additionally, each annotation line
+detail in Section \ref{featannot}. They can be hidden by toggling the {\sl Annotations
+$\Rightarrow$ Show annotations} menu option. Additionally, each annotation line
can be hidden and revealed in the same way as sequences {\sl via} the
pop-up context menu on the annotation name panel (Figure \ref{annot}).
Annotations can be reordered by dragging the annotation line label on the annotation label panel. Placing the
mouse over the top annotation label brings up a resize icon on the left. When this is
-displayed, Click-dragging up and down provides more space in the alignment window for viewing the annotations, and less space for the sequence alignment.
+displayed, click-dragging up and down provides either more space in the alignment window for viewing the annotations, or
+less space for the sequence alignment.
\begin{figure}
\begin{center}
\includegraphics[width=2.5in]{images/annot1.pdf}
\includegraphics[width=2.5in]{images/annot2.pdf}
\caption{{\bf Hiding Annotations} Annotations can either be hidden from the {\sl
-View} menu (left) or individually from the context menu (right).}
+Annotations} menu (left) or individually from the context menu opened by right clicking their label (right).}
\label{annot}
\end{center}
\end{figure}
-\exercise{Alignment Layout}{
-\label{exscreen}
-\exstep{Start Jalview and open the URL \textsf{http://www.jalview.org/examples/exampleFile.jar}.
-Select {\sl Format $\Rightarrow$ Wrap} from the alignment window menu.
-Experiment with the various options from the {\sl Format} menu, for example adjust the ruler placement,
-sequence ID format and so on. }
-\exstep{Hide all the annotation rows by selecting {\sl Annotations $\Rightarrow$
-Show Annotations} from the alignment window menu. Reveal the annotations by selecting the same menu option.} \exstep{Deselect {\sl Format $\Rightarrow$ Wrap}. Right click on the
-annotation row labels to bring up the pop-up context menu, then select {\sl
-Hide This Row}. Bring up the pop-up context menu again and select {\sl
-Show All Hidden Rows} to reveal them.}
-\exstep{Annotations can be reordered by clicking and dragging the row to the desired position. Click on the {\sl Consensus} row and drag it upwards to just above {\sl Quality}. The rows should now be reordered. Features and annotations are covered in more detail in Section \ref{featannot} below.}
-\exstep{Move the mouse to the top left hand corner of the Secondary Structure annotation row label -
-a grey up/down arrow symbol should appear - when this is shown, the height of the {\sl Annotation Area} can be changed
-by clicking and dragging the mouse up or down.}
-}
+%%TODO: multiple views - simple edits - observe changes in other views.
+
\subsection{Graphical Output}
\label{figuregen}
-\parbox[c]{4in}{Jalview allows alignments figures to be exported in three different formats, each of which is suited to a particular purpose. Image export is {\sl via} the {\sl File $\Rightarrow$ Export Image $\Rightarrow$ \ldots } alignment window menu option. }
-\parbox[c]{2in}{\centerline{\includegraphics[width=1.25in]{images/image.pdf}}}
+\parbox[c]{4in}{Jalview allows alignments figures to be exported in different formats, each of which is suited to a particular purpose.
+Image export is {\sl via} the {\sl File $\Rightarrow$ Export Image $\Rightarrow$ \ldots } alignment window menu option. }
+\parbox[c]{2in}{\centerline{\includegraphics[width=2in]{images/image.pdf}}}
\subsubsection{HTML}
-\parbox[c]{3in}{HTML is the format used by web pages. Jalview outputs the alignment as an HTML table with all the colours and fonts as seen. Any additional annotation will also be embedded as sensitive areas on the page, such as URL links for each sequence's ID label. This file can then be viewed directly with any web browser. Each residue is placed in an individual table cell. Unwrapped alignments will produce a very wide page.}
+\parbox[c]{4in}{HTML is the format used by web pages. Jalview outputs the
+alignment as a 'Scalable Vector Graphics' (or SVG) file with all the colours and fonts as seen, which is in turn embedded as a
+scrollable component within an HTML page.
+%% Functionality lost in 2.9 Any additional annotation will also be embedded as sensitive areas on the page, such as URL links for each sequence's ID label.
+This file can then be viewed directly with any web browser. Unwrapped alignments will produce a very wide page. Export options allow
+original data to be embedded in the HTML file as BioJSON.\footnote{BioJSON was introduced in Jalview 2.9 and fully described
+at \url{https://jalview.github.io/biojson/}}}
\parbox[c]{3.5in}{\centerline{\includegraphics[width=3in]{images/image_html.pdf}}}
\subsubsection{EPS}
-\parbox[c]{3in}{EPS is Encapsulated Postscript. {\bf It is the format of choice
+\parbox[c]{4in}{EPS is Encapsulated Postscript. {\bf It is the format of choice
for publications and posters} as it gives the highest quality output of any of
the image types. It can be scaled to any size, so will still look good on an A0
poster.
This format can be read by most good presentation and graphics packages such as Adobe Illustrator or Inkscape.
}
-\parbox[c]{3.5in}{\centerline{\includegraphics[width=3in]{images/image_eps.pdf}} \par \centerline{Zoom Detail of EPS image.}}
+\parbox[c]{4in}{\centerline{\includegraphics[width=2in]{images/image_eps.pdf}}
+\par \centerline{Zoom Detail of EPS image.}}
\subsubsection{PNG}
-\parbox[c]{3in}{
-PNG is Portable Network Graphics. This output option produces an image that can be easily included in web pages and incorporated in presentations using e.g. Powerpoint or Open Office. It is a bitmap image so does not scale and is unsuitable for use on posters, or in publications.
+\parbox[c]{4in}{
+PNG is Portable Network Graphics. This output option produces an image that can be easily included in web pages and incorporated in
+presentations using e.g. Powerpoint or Open Office. It is a bitmap image so does not scale and is unsuitable for use on posters,
+or in publications.
For submission of alignment figures to journals, please use EPS\footnote{If the journal complains, {\em insist}.}.
}
-\parbox[c]{3.5in}{\centerline{\includegraphics[width=3in]{images/image_png.pdf}} \par \centerline{Zoom Detail of PNG image.}}
+\parbox[c]{4in}{\centerline{\includegraphics[width=2in]{images/image_png.pdf}}
+\par \centerline{Zoom Detail of PNG image.}}
+
\exercise{Graphical Output}{
+ \label{graphicex}
\exstep{Load the example Jalview Jar file in Exercise \ref{exscreen}.
Customise it how you wish but leave it unwrapped.
Select {\sl File $\Rightarrow$ Export Image $\Rightarrow$ HTML} from the alignment menu.
alignment window but {\bf annotations are not exported}).}
\exstep{Export the alignment using the {\sl File $\Rightarrow$ Export Image $\Rightarrow$ PNG} menu option.
Open the file in an image viewer that allows zooming such as Paint or Photoshop (Windows), or Preview (Mac OS X) and zoom in.
-Notice that the image is a bitmap and it becomes pixelated very quickly.
+Notice that the image is a bitmap and it becomes pixelated when zoomed.
(Note that the {\bf annotation lines are included} in the image.)}
\exstep{Export the alignment using the {\sl File $\Rightarrow$ Export Image
$\Rightarrow$ EPS} menu option. Open the file in a suitable program such as
Photoshop, Illustrator, Inkscape, Ghostview, Powerpoint (Windows), or
Preview (Mac OS X). Zoom in and note that the image has near-infinite
-resolution.}
+resolution.}
+\exstep{Experiment with Jalview's other output options: try exporting an alignment view as 'BioJS', which employs the BioJS Multiple Sequence Alignment viewer. When would you use this type of export option ?}
+\exstep{Working with embedded BioJSON data. Drag and drop (or load via the file browser) the 'BioJS' HTML file in the previous step. Compare the original and imported alignment views - are there differences ?}
+\bf See the video at:
+\url{http://www.jalview.org/training/Training-Videos}.
}
-The next chapters introduce Jalviews analysis features. Chapter \ref{featannot}
-describes the mechanisms provided by Jalview for interactive creation of sequence and alignment annotation, and how they can be displayed, imported and exported and used to reorder the alignment. Chapter
-\ref{featuresfromdb} discusses the retrieval of database references and
-establishment of sequence coordinate systems for the retrieval and display of
-features from databases and DAS annotation services.
-Chapter \ref{msaservices} describes how to use the range of multiple alignment
-programs provided by JABAWS, and Chapter \ref{aacons} introduces JABAWS AACon
-service for protein multiple alignment conservation analysis.
- In Chapter \ref{alignanalysis}, you will find
-descriptions and exercises on building and displaying trees, PCA analysis,
-alignment redundancy removal, pairwise alignments and alignment conservation
-analysis.
-Chapter \ref{wkwithstructure} introduces the structure visualization
-capabilities of Jalview.
-Chapter \ref{protsspredservices} explains how to perform protein secondary
-structure predictions with JPred, and JABAWS protein disorder prediction
-services are introduced in Chapter \ref{protdisorderpred}.
-Chapter \ref{workingwithnuc} describes functions and visualization techniques relevant
-to working with nucleotide sequences, coding region annotation and nucleotide
-sequence alignments.
-Chapter \ref{jvwebservices} introduces the various web based services
-available to Jalview users, and Chapter \ref{jabaservices} explains how to
-configure the Jalview Desktop for access to new JABAWS servers.
+% left out for Glasgow 2016
+% \newpage
+%
+% \section{Summary - the rest of the manual}
+%
+% The first few chapters have covered the basics of Jalview operation: from
+% starting the program, importing, exporting, selecting, editing and colouring
+% aligments, to the generation of figures for publication, presentation and web
+% pages.
+%
+% The remaining chapters in the manual cover:
+%
+% \begin{list}{$\circ$}{}
+% \item{Chapter \ref{featannot} covers the creation, manipulation and visualisation
+% of sequence and alignment annotation, and retrieval of sequence and feature data
+% from databases.}
+% \item {Chapter \ref{msaservices} explores the range of multiple alignment
+% programs offered via Jalview's web services, and introduces the use of
+% AACon for protein multiple alignment conservation analysis.}
+% \item {Chapter \ref{alignanalysis} introduces Jalview's built in tools for
+% multiple sequence alignment analysis, including trees, PCA, and alignment
+% conservation analysis. }
+% \item {Chapter \ref{3Dstructure} demonstrates the structure visualization
+% capabilities of Jalview.}
+% \item {Chapter \ref{proteinprediction} introduces protein sequence based
+% secondary structure and disorder prediction tools, including JPred.}
+% \item {Chapter \ref{dnarna} covers the special functions and
+% visualization techniques for working with RNA alignments and protein coding
+% sequences.}
+% \item {Chapter \ref{jvwebservices} provides instructions on the
+% installation of your own Jalview web services.}
+% \end{list}
\chapter{Annotation and Features}
\label{featannot}
overlaid on the sequences and the alignment.
Generally speaking, annotations reflect properties of the alignment as a
whole, often associated
-with columns in the alignment. Whilst features are associated with specific residues in the sequence.
+with columns in the alignment. Features are often associated with specific
+residues in the sequence.
Annotations are shown below the alignment in the annotation panel, the
properties are often based on the alignment.
but are shown mapped on to specific residues within the alignment.
Features and annotation can be interactively created, or retrieved from external
-data sources. DAS (the Distributed Annotation System) is the primary source of
-sequence features, whilst webservices like JNet (see \ref{jpred} above) can be used to analyse a
-given sequence or alignment and generate annotation for it.
+data sources. Webservices like JPred (see \ref{jpred}) can be used to
+analyse a given sequence or alignment and generate annotation for it.
-\section{Conservation, Quality and Conservation Annotation}
+\section{Conservation, Quality, Consensus and other Annotation}
\label{annotationintro}
Jalview automatically calculates several quantitative alignment annotations
which are displayed as histograms below the multiple sequence alignment columns.
-Conservation, quality and conservation scores are examples of dynamic
+Conservation, quality and consensus scores are examples of dynamic
annotation, so as the alignment changes, they change along with it.
The scores can be used in the hybrid colouring options to shade the alignments.
Mousing over a conservation histogram reveals a tooltip with more information.
for all mutations, the ratio of the two BLOSUM 62 scores for a mutation pair and each residue's conserved BLOSUM62 score (which is higher).
This value is normalised for each column, and then plotted on a scale from 0 to 1.
+\subsubsection{Occupancy Annotation}
+
+Alignment occupancy simply reflects the number of residues aligned at each column
+in the multiple sequence alignment. To see this annotation you may first need to enable it by ticking the {\sl Occupancy} check-box
+in the {\sl Visual} tab in Jalview's {\sl Preferences} before opening an alignment. Occupancy is particularly useful in conjunction with
+the {\sl Select/Hide by Annotations} dialog since it allows the view to be filtered to exclude regions of the alignment with a high proportion of gaps.
+
\subsubsection{Group Associated Annotation}
\label{groupassocannotation}
Group associated consensus and conservation annotation rows reflect the
\subsection{Creating User Defined Annotation}
To create a new annotation row, right click on the annotation label panel and select the {\sl Add New Row} menu option (Figure \ref{newannotrow}).
-A dialogue box appears. Enter the label to use for this row and a new row will appear.
+A dialog box appears. Enter the label to use for this row and a new row will appear.
To create a new annotation, first select all the positions to be annotated on the appropriate row.
Right-clicking on this selection brings up the context menu which allows the insertion of graphics for secondary structure ({\sl Helix} or {\sl Sheet}),
-text {\sl Label} and the colour in which to present the annotation (Figure \ref{newannot}). On selecting {\sl Label} a dialogue box will appear,
+text {\sl Label} and the colour in which to present the annotation (Figure \ref{newannot}). On selecting {\sl Label} a dialog box will appear,
requesting the text to place at that position. After the text is entered, the selection can be removed and the annotation becomes clearly
visible\footnote{When annotating a block of positions, the text can be partly obscured by the selection highlight. Pressing the [ESC] key clears
the selection and the label is then visible.}. Annotations can be coloured or deleted as desired.
G.J. (1993) {\sl CABIOS } {\bf 9}, 745-756}.
Consensus is the modal residue (or {\tt +} where there is an equal top residue).
The inclusion of gaps in the consensus calculation can be toggled by
-right-clicking on the the Consensus label and selecting {\sl Ignore Gaps in
+right-clicking on the Consensus label and selecting {\sl Ignore Gaps in
Consensus} from the pop-up context menu located with consensus annotation row.
Quality is a measure of the inverse likelihood of unfavourable mutations in the alignment. Further details on these
calculations can be found in the on-line documentation.
\exercise{Annotating Alignments}{
+ \label{annotatingalignex}
\exstep{Load the alignment at \textsf{http://www.jalview.org/tutorial/alignment.fa}.
-Right-click on the {\sl Conservation} annotation row to
-bring up the context menu and select {\sl Add New Row}. A dialogue box will appear asking for {\sl Label for annotation}.
-Enter ``Iron binding site" and click OK. A new, empty, row appears.
+Right-click on the label name of the {\sl Conservation} annotation row to
+bring up the context menu and select {\sl Add New Row}. A dialog box will
+appear asking for {\sl Annotation Name} and {\sl Annotation Description}.
+Enter "Iron binding site" and click {\sl OK}. A new, empty, row appears.
}
\exstep{
Navigate to column 97. Move down and on the new annotation row called
-``Iron binding site, select column 97.
+"Iron binding site", select column 97.
Right click at this selection and select {\sl Label} from the context menu.
-Enter ``Fe" in the box and click OK. Right-click on the selection again and select {\sl Colour}.
-Choose a colour from the colour chooser dialogue
-and click OK. Press [ESC] to remove the selection.
+Enter "Fe" in the box and click {\sl OK}. Right-click on the selection again
+and select {\sl Colour}.
+Choose a colour from the colour chooser dialog
+and click {\sl OK}. Press [ESC] to remove the selection.
+
+{\sl Note: depending on your Annotation sort settings, your newly
+created annotation row might "jump" to the top or bottom of the annotation
+panel. Just scroll up or down to find it again - the column you marked will
+still be selected. }
+
}
\exstep{ Select columns 70-77 on the annotation row. Right-click and choose {\sl Sheet} from the
- context menu. You will be prompted for a label. Enter ``B" and press OK. A new line showing the
+ context menu. You will be prompted for a label. Enter "B" and press {\sl OK}. A new line showing the
sheet as an arrow appears. The colour of the label can be changed but not the colour of the sheet
arrow.
}
-\exstep{Right click on the title text of annotation row that you just created.
-Select {\sl Export Annotation} and, in the {\bf Export Annotation} dialog box that will open, select the Jalview format and click
-the [To Textbox] button.
-
-The format for this file is given in the Jalview help. Press [F1] to open it, and find
-the ``Annotations File Format'' entry in the ``Alignment Annotations'' section of the contents
-pane. }
-
-\exstep{Export the file to a text editor and edit the file to change the name of the annotation
-row. Save the file and drag it onto the alignment view.}
-\exstep{Try to add an additional helix somewhere along the row by editing the file and
-re-importing it.
-{\sl Hint: Use the {\bf Export Annotation} function to view what helix annotation looks like in
+\exstep{Right click on the title text in the annotation row that you just
+created.
+Select {\sl Export Annotation} in context menu and, in the Export Annotation
+dialog box that will open, select the Jalview format and click the {\sl [To Textbox]} button.
+(The format for this file is given in the Jalview help. Press [F1] to open it ([F1]-[Fn] (Mac)),
+and find the "Annotations File Format" entry in the "Alignment Annotations" section of the contents
+pane.) }
+
+\exstep{Open a text editor and copy the annotation text into the editor.
+Edit the text by changing the name of the annotation row and save the file.}
+\exstep{Drag the file onto the alignment in Jalview and check the annotation
+panel.} \exstep{Return to the text editor, add an additional helix somewhere
+along the row, save the file and re-importing it into Jalview as previously.
+{\sl Hint: Use the Export Annotation function to view what helix annotation looks like in
a Jalview annotation file.}}
-\exstep{Use the {\sl Alignment Window $\Rightarrow$ File $\Rightarrow$ Export Annotations...}
-function to export all the alignment's annotation to a file.}
-\exstep{Open the exported annotation in a text editor, and use the {\bf Annotation File Format}
+\exstep{In the alignment window menu, select {\sl File $\Rightarrow$ Export Annotations...}
+to export all the alignment's annotation to a file. Save the file.}
+\exstep{Open the exported annotation in a text editor, and use the Annotation File Format
documentation to modify the style of the Conservation, Consensus and Quality annotation rows so
they appear as several lines on a single line graph.
{\sl Hint: You need to change the style of annotation row in the first field of the annotation
-row entry in the file, and create an annotation row grouping to overlay the three quantitative
-annotation rows.}
+row entry in the file. Create an annotation row grouping to overlay the
+three quantitative annotation rows.}
+}
+\exstep{{\bf Homework once you have completed exercise
+\ref{secstrpredex}:}
+\label{viewannotfileex}
+
+Recover or recreate the secondary structure predictions that you made from
+JPred. Use the {\sl File $\Rightarrow$ Export Annotation} function to view the Jnet secondary structure prediction annotation row.
+Note: the
+SEQUENCE\_REF statements surrounding the row specifying the sequence association for the
+annotation.
}
-\label{viewannotfileex}\exstep{Recover or recreate the secondary structure
-prediction that you made in exercise \ref{secstrpredex}. Use the {\sl File $\Rightarrow$ Export
-Annotation} function to view the Jnet secondary structure prediction annotation row. Note the
-{\bf SEQUENCE\_REF} statements surrounding the row specifying the sequence association for the
-annotation. } }
+\bf See the video at:
+\url{http://www.jalview.org/training/Training-Videos}.}
+
+
+\section{Sequence Features}
+Sequence features are annotation associated with a specific sequence - often marking a specific region, such as a domain or binding site. Jalview allows features to be created simply by selecting the area in a sequence (or sequences) to form the feature and selecting {\sl Selection $\Rightarrow$ Create Sequence Feature } from the right-click context menu (Figure \ref{features}). A dialog box allows the user to customise the feature with respect to name, group, and colour. The feature is then associated with the sequence. Moving the mouse over a residue associated with a feature brings up a tool tip listing all features associated with the residue.
+
+\begin{figure}[htbp]
+\begin{center}
+\includegraphics[width=2in]{images/feature1.pdf}
+\includegraphics[width=2.5in]{images/feature2.pdf}
+\includegraphics[width=1.5in]{images/feature3.pdf}
+\caption{{\bf Creating sequence features.} Features can readily be created from selections via the context menu and are then displayed on the sequence. }
+\label{features}
+\end{center}
+\end{figure}
+Creation of features from a selection spanning multiple sequences results in the creation of one feature per sequence.
+Each feature remains associated with its own sequence.
+
+%%TODO: change EMBL to ENA in Jalview and the Manual !
\section{Importing Features from Databases}
\label{featuresfromdb}
-Jalview supports feature retrieval from public databases either directly or {\sl
-via} the Distributed Annotation System (DAS\footnote{http://www.biodas.org/}).
-It includes built in parsers for Uniprot and ENA (or EMBL) records retrieved
+Jalview supports feature retrieval from public databases.
+It includes built in parsers for Uniprot, Ensembl and ENA (or EMBL) records retrieved
from the EBI. Sequences retrieved from these sources using the sequence fetcher (see
Section \ref{fetchseq}) will already possess features.
+
+\begin{figure}[htbp]
+\begin{center}
+\includegraphics[width=4in]{images/features4.pdf}
+\caption{{\bf Multiple sequence features.} An alignment with JPred secondary structure prediction annotation below it, and many sequence features overlaid onto the aligned sequences. The tooltip lists the features annotating the residue below the mouse-pointer.}
+\end{center}
+\end{figure}
+
+
\subsection{Sequence Database Reference Retrieval}
\label{fetchdbrefs}
Jalview maintains a list of external database references for each sequence in
You can export all the database cross references and annotation terms shown in
the sequence ID tooltip for a sequence by right-clicking and selecting the {\sl
-[Sequence ID] $\Rightarrow$ Sequence details \ldots} option from the popup menu.
+[Sequence ID] $\Rightarrow$ Sequence Details} option from the popup
+menu.
A similar option is provided in the {\sl Selection} sub-menu allowing you to
obtain annotation for the sequences currently selected.
Jalview includes a function to automatically verify and update each sequence's
start and end numbering against any of the sequence databases that the {\sl
Sequence Fetcher} has access to. This function is accessed from the {\sl
-Webservice $\Rightarrow$ Fetch DB References} sub-menu in the Alignment
+Web Service $\Rightarrow$ Fetch DB References} sub-menu in the Alignment
window. This menu allows you to query either the set of {\sl Standard
-Databases}, which includes EMBL, Uniprot, the PDB, and the currently selected
-DAS sequence sources, or just a specific datasource from one of the submenus.
+Databases}, which includes EMBL, Ensembl, Uniprot, the PDB, or just a specific datasource from one of the submenus.
When one of the entries from this menu is selected, Jalview will use the ID
string from each sequence in the alignment or in the currently selected set to
retrieve records from the external source. Any sequences that are retrieved are
the database did not exactly contain the sequence given in the alignment (the
{\sl ``Sequence not 100\% match'' dialog box}).
-\exercise{Retrieving Database References}{
-\exstep{Load the example alignment at http://www.jalview.org/tutorial/alignment.fa}
-\exstep{Verify that there are no database references for the sequences by first
-checking that the {\sl View $\Rightarrow$ Sequence ID Tooltip $\Rightarrow$ Show
-Database Refs} option is selected, and then mousing over each sequence's ID.}
-\exstep{Use the {\sl Webservice $\Rightarrow$ Fetch DB References} menu option to retrieve database IDs for the sequences.}
-\exstep{Examine the tooltips for each sequence in the alignment as the retrieval progresses - note the appearance of new database references.}
-\exstep{Once the process has finished, save the alignment as a Jalview Project.}
-\exstep{Now close all the windows and open the project again, and verify that the database references and sequence features are still present on the alignment}
-\exstep{View the {\sl Sequence details \ldots} report for the FER1\_SPIOL sequence and for the whole alignment. Which sequences have web links associated with them ?}
+\subsubsection{Rate of Feature Retrieval}
+Feature retrieval can take some time if a large number of sources are selected
+and if the alignment contains a large number of sequences.
+As features are retrieved, they are immediately added to the current alignment view.
+The retrieved features are shown on the sequence and can be customised as described previously.
-}
-\subsection{Retrieving Features {\sl via} DAS}
-\label{dasfretrieval}
-Jalview includes a client to retrieve features from DAS annotation servers. To
-retrieve features, select {\sl View $\Rightarrow$ Feature Settings\ldots} from the alignment window menu. Select the {\sl DAS Settings} tab in the Sequence Feature Settings Window (Figure \ref{das}). A list of DAS sources compiled from the currently configured DAS registry\footnote{By default, this will be the major public DAS server registry maintained by the Sanger Institute: http://www.dasregistry.org} is shown in the left hand pane. Highlighting an entry on the left brings up information about that source in the right hand panel.
+\subsection{Customising Feature Display}
\begin{figure}[htbp]
\begin{center}
-\includegraphics[width=2.5in]{images/das1.pdf}
-\includegraphics[width=2.5in]{images/das2.pdf}
-\caption{{\bf Retrieving DAS annotations.} DAS features are retrieved using the {\sl DAS Settings} tab (left) and their display customised using the {\sl Feature Settings} tab (right).}
-\label{das}
+\includegraphics[width=5in]{images/features5.pdf}
+\caption{{\bf Customising sequence features.} Features can be recoloured, switched on or off and have the rendering order changed. }
+\label{custfeat}
\end{center}
\end{figure}
-Select appropriate DAS sources as required then click on {\sl Fetch DAS
-Features}. If you know of additional sources not listed in the configured
-registry, then you may add them with the {\sl Add Local Source} button. Use
-the {\sl Authority},{\sl Type}, and {\sl Label} filters to restrict the list
-of sources to just those that will return features for the sequences in the
-alignment.
-
-Following DAS feature retrieval, the {\sl Feature Settings} panel takes on a
-slightly different appearance (Figure \ref{das} (right)). Each data source is
-listed and groups of features from one data source can be selected/deselected
-by checking the labelled box at the top of the panel.
+Feature display can be toggled on or off by selecting the {\sl View
+$\Rightarrow$ Show Sequence Features} menu option. When multiple features are
+present it is usually necessary to customise the display. Jalview allows the
+display, colour, rendering order and transparency of features to be modified
+{\sl via} the {\sl View $\Rightarrow$ Feature Settings\ldots} menu option. This
+brings up a dialog window (Figure \ref{custfeat}) which allows the
+visibility of individual feature types to be selected, assigned colours to be changed (by
+clicking on the colour of each sequence feature type) and the rendering order
+modified by dragging feature types to a new position in the list. Dragging the
+slider alters the transparency of the feature rendering. Clicking in the {\sl Configuration} column
+ opens the {\sl Display Settings} dialog which allows more complex shading schemes
+ and also the creation of filters, and right-clicking opens a context sensitive menu
+ that offers options for selecting and hiding columns or sorting the alignment according the feature's distribution or score attribute.
+These capabilities are described further in sections
+\ref{featureschemes} and \ref{featureordering}.
-\exercise{Retrieving Features with DAS}{
-\label{dasfeatretrexcercise}
-\exstep{Load the alignment at
-\textsf{http://www.jalview.org/tutorial/alignment.fa}. Select {\sl View
-$\Rightarrow$ Feature Settings \ldots} from the alignment window menu. Select
-the {\sl DAS Settings} tab. A long list of available DAS sources is listed.
-Select a small number, eg Uniprot, DSSP, signalP and netnglyc. Click.
-A window may prompt whether you wish Jalview to fetch DAS features. Click {\sl
-Yes}.
-Jalview will start retrieving features. As features become available they will be mapped onto the alignment. }
-\exstep{If Jalview is taking too long to retrieve features, the process can be cancelled with the {\sl Cancel Fetch} button.
-Rolling the mouse cursor over the sequences reveals a large number of features annotated in the tool tip.
-Close the Sequence Feature Settings window. }
-\exstep{Move the mouse over the sequence ID panel.
-Non-positional features such as literature references and protein localisation predictions are given in the tooltip, below any database cross references associated with the sequence.}
-\exstep{Search through the alignment to find a feature with a link symbol next to it.
-Right click to bring up the alignment view popup menu, and find a corresponding entry in the {\sl Link } sub menu. }
-% TODO this doesn't work ! \includegraphics[width=.3in]{images/link.pdf}
-\exstep{
-Select {\sl View $\Rightarrow$ Feature Settings\ldots} to reopen the Feature Settings window. All the loaded feature types should now be displayed. Those at the top of the list are drawn on top of those below, obscuring them in the alignment view where they overlap. Move the feature settings window so that the alignment is visible and uncheck some of the feature types by clicking the tick box in the display column. Observe how the alignment display changes. Note that unselected feature types do not appear in the tool tip.
-}
-\exstep{Reorder the features by dragging feature types up and down the order in the Feature Settings panel. e.g. Click on {\sl CHAIN} then move the mouse downwards to drag it below {\sl DOMAIN}. Note that {\sl DOMAIN} is now shown on top of {\sl CHAIN} in the alignment window. Drag {\sl METAL} to the top of the list. Observe how the cysteine residues are now highlighted as they have a {\sl METAL} feature associated with them.
-}
-
-\exstep{Press the {\sl Optimise Order} button. The features will be ordered according to increasing length, placing features that annotate shorter regions of sequence higher on the display stack.}
-
-\exstep{Select {\sl File $\Rightarrow$ Export Features\ldots} from the Alignment window. You can choose to export the retrieved features as a GFF file, or Jalview's own Features format.
-% TODO: describe working with features files and GFF
-}
-}
-
-\subsubsection{The Fetch Uniprot IDs Dialog Box}
-\label{discoveruniprotids}
-If any sources are selected which refer to Uniprot coordinates as their reference system,
-then you may be asked if you wish to retrieve Uniprot IDs for your sequence. Pressing OK instructs Jalview to verify the sequences against Uniprot records retrieved using the sequence's ID string. This operates in much the same way as the {\sl Web Service $\Rightarrow$ Fetch Database References } function described in Section \ref{fetchdbrefs}. If a sequence is verified, then the start/end numbering will be adjusted to match the Uniprot record to ensure that features retrieved from the DAS source are rendered at the correct position.
-
-\subsubsection{Rate of Feature Retrieval}
-Feature retrieval can take some time if a large number of sources is selected and if the alignment
-contains a large number of sequences. This is because Jalview only queries a particular DAS source with one sequence at a time, to avoid overloading it. As features are retrieved, they are immediately added to the current alignment view. The retrieved features are shown on the sequence and can be customised as described previously.
-
-
-\subsection{Colouring Features by Score or Description
-Text}
+\subsection{Changing how Features are coloured and displayed}
\label{featureschemes}
Sometimes, you may need to visualize the differences in information carried by
sequence features of the same type. This is most often the case when features
-of a particular type are the result of a specific type of database query or calculation. Here, they may also carry information within their textual description, or most commonly for calculations, a score related to the property being investigated. Jalview can shade sequence
-features using a graduated colourscheme in order to highlight these variations.
-In order to apply a graduated scheme to a feature type, select the `Graduated
-colour' entry in the Sequence {\sl Feature Type}'s popup menu, which is opened by
-right-clicking the {\sl Feature Type} or {\sl Color} in the {\sl Sequence Feature Settings} dialog box. Two types
-of colouring styles are currently supported: the default is quantitative
-colouring, which shades each feature based on its score, with the highest
-scores receiving the `Max' colour, and the lowest scoring features coloured
-with the `Min' colour. Alternately, you can select the `Colour by label'
+of a particular type are the result of a specific type of database query or calculation.
+Here, they may also carry information within their textual description, or most commonly
+for calculations, a score related to the property being investigated. Features imported
+from genomic databases and Variant Call Format files may also have a number of additional
+attributes. Jalview allow filters and different types of colouring to be applied to allow variations in these attributes to be highlighted.
+In order to create a filter or modify the way a feature type is coloured, select the `Configuration' column for that {\sl Feature Type} in the {\sl Sequence Feature Settings} dialog box.
+
+Instead of shading a feature with an assigned colour according to its type, you can select the `Colour by text'
option to create feature colours according to the description text associated
with each feature. This is useful for general feature types - such as
Uniprot's `DOMAIN' feature - where the actual type of domain is given in the
-feature's description.
+feature's description. If other attributes are present you can choose one of them from the drop-down menu.
-Graduated feature colourschemes can also be used to exclude low or
+If Scores or numeric attributes are present, the {\sl Graduated Colour} section of the dialog allows a quantitative
+shading scheme to be defined, with the highest
+scores receiving the `Max' colour, and the lowest scoring features coloured
+with the `Min' colour. Alternately,
+you can define a threshold to exclude low or
high-scoring features from the alignment display. This is done by choosing your
desired threshold type (either above or below), using the drop-down menu in the
dialog box. Then, adjust the slider or enter a value in the text box to set the
threshold for displaying this type of feature.
-The feature settings dialog box allows you to toggle between a graduated and
-simple feature colourscheme using the pop-up menu for the feature type. When a
-graduated scheme is applied, it will be indicated in the colour column for
-that feature type - with coloured blocks or text to indicate the colouring
-style and a greater than ($>$) or less than ($<$) symbol to indicate when a
-threshold has been defined.
+When a filters and complex colourschemes are applied, the configuration column will show coloured blocks or text to indicate the colouring
+style and any attribute filters.
+
\subsection{Using Features to Re-order the Alignment}
\label{featureordering}
The presence of sequence features on certain sequences or in a particular
region of an alignment can quantitatively identify important trends in
the aligned sequences. In this case, it is more useful to
-re-order the alignment based on the number of features or their associated scores, rather than simply re-colour the aligned sequences. The sequence feature settings
-dialog box provides two buttons: `Seq sort by Density' and `Seq sort by
+re-order the alignment based on the number of features or their associated scores, rather than simply re-colour the aligned sequences.
+The sequence feature settings
+dialog box provides two buttons: `Sequence sort by Density' and `Sequence sort by
Score', that allow you to reorder the alignment according to the number of
sequence features present on each sequence, and also according to any scores
associated with a feature. Each of these buttons uses the currently displayed
% colour styles are encoded. }
% }
-\subsection{Creating Sequence Features}
-Sequence features can be created simply by selecting the area in a sequence (or sequences) to form the feature and selecting {\sl Selection $\Rightarrow$ Create Sequence Feature } from the right-click context menu (Figure \ref{features}). A dialogue box allows the user to customise the feature with respect to name, group, and colour. The feature is then associated with the sequence. Moving the mouse over a residue associated with a feature brings up a tool tip listing all features associated with the residue.
-
-\begin{figure}[htbp]
-\begin{center}
-\includegraphics[width=2in]{images/feature1.pdf}
-\includegraphics[width=2.5in]{images/feature2.pdf}
-\includegraphics[width=1.5in]{images/feature3.pdf}
-\caption{{\bf Creating sequence features.} Features can readily be created from selections via the context menu and are then displayed on the sequence. }
-\label{features}
-\end{center}
-\end{figure}
-
-Creation of features from a selection spanning multiple sequences results in the creation of one feature per sequence.
-Each feature remains associated with its own sequence.
-
-\subsection{Customising Feature Display}
-
-Feature display can be toggled on or off by selecting the {\sl View
-$\Rightarrow$ Show Sequence Features} menu option. When multiple features are
-present it is usually necessary to customise the display. Jalview allows the
-display, colour, rendering order and transparency of features to be modified
-{\sl via} the {\sl View $\Rightarrow$ Feature Settings\ldots} menu option. This
-brings up a dialogue window (Figure \ref{custfeat}) which allows the
-visibility of individual feature types to be selected, colours changed (by
-clicking on the colour of each sequence feature type) and the rendering order
-modified by dragging feature types to a new position in the list. Dragging the
-slider alters the transparency of the feature rendering. The Feature
-Settings dialog also includes functions for more advanced feature shading
-schemes and buttons for sorting the alignment according to the distribution of
-features. These capabilities are described further in sections
-\ref{featureschemes} and \ref{featureordering}.
-
-\begin{figure}[htbp]
-\begin{center}
-\includegraphics[width=4in]{images/features4.pdf}
-\caption{{\bf Multiple sequence features.} An alignment with JPred secondary structure prediction annotation below it, and many sequence features overlaid onto the aligned sequences. The tooltip lists the features annotating the residue below the mouse-pointer.}
-\end{center}
-\end{figure}
-
-\begin{figure}[htbp]
-\begin{center}
-\includegraphics[width=4in]{images/features5.pdf}
-\caption{{\bf Customising sequence features.} Features can be recoloured, switched on or off and have the rendering order changed. }
-\label{custfeat}
-\end{center}
-\end{figure}
\subsection{Sequence Feature File Formats}
features file.
\exercise{Creating Features}{
+\label{featuresex}
\exstep{Open the alignment at \textsf{http://www.jalview.org/tutorial/alignment.fa}.
We know that the Cysteine residues at columns 97, 102, 105 and 135 are involved in
iron binding so we will create them as features. Navigate to column 97, sequence 1.
Select the entire column by clicking in the ruler bar. Then right-click on the selection
to bring up the context menu and select {\sl Selection $\Rightarrow$ Create Sequence Feature}.
-A dialogue box will appear.
+A dialog box will appear.
}
\exstep{
Enter a suitable Sequence Feature Name (e.g. ``Iron binding site") in the
appropriate box. Click on the Feature Colour bar to change the colour if
-desired, add a short description (``One of four Iron binding Cysteines") and press OK. The features will then appear on the sequences. } \exstep{Roll the mouse cursor over the new features. Note that the position given in the tool tip is the residue number, not the column number. To demonstrate that there is one feature per sequence, clear all selections by pressing [ESC] then insert a gap in sequence 3 at column 95. Roll the mouse over the features and you will see that the feature has moved with the sequence. Delete the gap you created.
+desired, add a short description (``One of four Iron binding Cysteines") and
+press {\sl OK}. The features will then appear on the sequences. } \exstep{Roll
+the mouse cursor over the new features.
+Note that the position given in the tool tip is the residue number, not the column number.
+To demonstrate that there is one feature per sequence, clear all selections by pressing [ESC],
+ then insert a gap in sequence 3 at column 95 using the [SHIFT] key.
+Roll the mouse over the features and you will see that the feature has moved
+with the sequence. Delete the gap you created using {\sl Edit
+$\Rightarrow$ Undo}.
}
\exstep{
-Add a similar feature to column 102. When the feature dialogue box appears, clicking the Sequence Feature
+Add a similar feature to column 102. When the feature dialog box appears, clicking the Sequence Feature
Name box brings up a list of previously described features. Using the same Sequence Feature Name allows the features to be grouped.}
\exstep{Select {\sl View $\Rightarrow$ Feature Settings\ldots} from the
alignment window menu. The Sequence Feature Settings window will appear. Move
this so that you can see the features you have just created. Click the check
box for ``Iron binding site" under {\sl Display} and note that display of this
feature type is now turned off. Click it again and note that the features are
-now displayed. Close the sequence feature settings box by clicking OK or
-Cancel.} }
+now displayed. Close the sequence feature settings box by clicking {\sl OK} or
+{\sl Cancel}.}
+
+\exstep{Select {\sl View $\Rightarrow$ Show Sequence Features} from the
+alignment window menu. The sequence features are now hidden. Repeat this step and the features are
+displayed.}
+
+\bf See the video at:
+\url{http://www.jalview.org/training/Training-Videos}.}
\chapter{Multiple Sequence Alignment}
\label{msaservices}
McWilliam H, Remmert M, Soding J, Thompson JD, Higgins DG (2011) {\sl Molecular
Systems Biology} {\bf 7} 539
\href{http://dx.doi.org/10.1038/msb.2011.75}{doi:10.1038/msb.2011.75}} Of these,
-T-COFFEE is slow but the accurate. ClustalW is historically
+T-COFFEE is slow but accurate. ClustalW is historically
the most widely used. Muscle is fast and probably best for
smaller alignments. MAFFT is probably the best for large alignments,
however Clustal Omega, released in 2011, is arguably the fastest and most
accurate tool for protein multiple alignment.
+\section{Performing a multiple sequence alignment}
To run an alignment web service, select the appropriate method from the {\sl
Web Service $\Rightarrow$ Alignment $\Rightarrow$ \ldots} submenu (Figure
\ref{webservices}). For each service you may either perform an alignment with
using the `Algorithm ordering' entry within the {\sl Calculate $\Rightarrow$
Sort } sub menu.
-\subsubsection{Realignment}
+\subsection{Realignment to add sequences to an existing alignment}
The re-alignment option is currently only supported by Clustal
Omega and ClustalW. When performing a re-alignment, Jalview submits the
-current selection to the alignment service complete with any existing gaps. This
-approach is useful when one wishes to align additional sequences to an existing alignment without
-any further optimisation to the existing alignment. The re-alignment service
-provided by ClustalW in this case is effectively a simple form of profile
-alignment.
+current selection to the alignment service complete with any existing gaps.
+Realignment with ClustalW is useful when one wishes to align
+additional sequences to an existing alignment without any further optimisation
+to the existing alignment. ClustalO's realignment works by generating a
+probabilistic model (a.k.a HMM) from the original alignment, and then realigns
+{\bf all} sequences to this profile. For a well aligned MSA, this process
+will simply reconstruct the original alignment (with additional sequences), but
+in the case of low quality MSAs, some differences may be introduced.
\begin{figure}[htbp]
\begin{center}
\end{center}
\end{figure}
-\subsubsection{Alignments of Sequences that include Hidden Regions}
-If the view or selected region that is submitted for alignment contains hidden
+\subsection{Alignments of Sequences that include Hidden Regions}
+If the view or selected region submitted for alignment contains hidden
regions, then {\bf only the visible sequences will be submitted to the service}.
Furthermore, each contiguous segment of sequences will be aligned independently
(resulting in a number of alignment `subjobs' appearing in the status window).
columns can be used to preserve existing parts of an alignment whilst the
visible parts are locally refined.
-
-\subsection{Customising the Parameters used for Alignment}
-JABA web services allow you to vary the parameters used when performing a
-bioinformatics analysis. For JABA alignment services, this means you are
-usually able to modify the following types of parameters:
-\begin{list}{$\bullet$}{}
-\item Amino acid or nucleotide substitution score matrix
-\item Gap opening and widening penalties
-\item Types of distance metric used to construct guide trees
-\item Number of rounds of re-alignment or alignment optimisation
-\end{list}
-
-\subsubsection{Getting Help on the Parameters for a Service}
-Each parameter available for a method usually has a short description, which
-Jalview will display as a tooltip, or as a text pane that can be opened under
-the parameter's controls. In the parameter shown in Figure
-\ref{clustalwparamdetail}, the description was opened by selecting the button on the left hand side. Online help for the
-service can also be accessed, by right clicking the button and selecting a URL
-from the pop-up menu that will open.
+\subsection{Alignment Service Limits}
+Multiple alignment is a computationally intensive calculation. Some JABA server
+services and service presets only allow a certain number of sequences to be
+aligned. The precise number will depend on the server that you are using to
+perform the alignment. Should you try to submit more sequences than a service
+can handle, then an error message will be shown informing you of the maximum
+number allowed by the server.
\exercise{Multiple Sequence Alignment}{
-\exstep{ Close all windows and open the alignment at {\sf
+\label{msaex}
+\exstep{ Close all windows. Open the alignment at {\sf
http://www.jalview.org/tutorial/unaligned.fa}. Select {\sl
Web Service $\Rightarrow$ Alignment $\Rightarrow$ Muscle with Defaults}.
A window will open giving the job status. After a short time, a second window will open
with the results of the alignment.}
\exstep{Return to the first sequence alignment window by clicking on
- the window, and repeat using ClustalO (Omega) and MAFFT (from the {\sl Web
- Service $\Rightarrow$ Alignment} menu) on the same initial alignment. Compare them and
- you should notice small differences. }
-\exstep{Select the last three sequences in the MAFFT alignment, and de-align them
+ the window, and repeat using {\sl ClustalO} (Omega) ({\sl with Defaults}), from the
+ {\sl Web Service $\Rightarrow$ Alignment} menu, using the same initial
+ alignment.
+ Compare them and you should notice small differences. }
+\exstep{Select the last three sequences in the MAFFT alignment (you may need the scroll down the alignment), and de-align them
with {\sl Edit $\Rightarrow$ Remove All Gaps}. Press [ESC] to deselect these
-sequecnes. Then submit this view for re-alignment with ClustalO.}
-\exstep{Return to the window in section (c), use [CTRL]-Z (undo) to recover the
-alignment of the last three sequences in this MAFFT alignment.
+sequences. Then submit this view for re-alignment with {\sl ClustalO}.}
+\exstep{Return to the alignment window in section (c), use [CTRL]-Z (undo) to
+recover the alignment of the last three sequences in this MAFFT alignment.
Once the ClustalO re-alignment has completed, compare the results of
re-alignment of the three sequences with their alignment in the original MAFFT result.}
\exstep{Select columns 60 to 125 in the original MAFFT alignment and hide them,
by right clicking the mouse to bring up context menu.
-Select {\sl Web Services $\Rightarrow$ Alignment $\Rightarrow$ Mafft with Defaults} to
+Select {\sl Web Service $\Rightarrow$ Alignment $\Rightarrow$ Mafft with Defaults} to
submit the visible portion of the alignment to MAFFT. When the web service job pane appears,
note that there are now two alignment job status panes shown in the window.}
\exstep{When the MAFFT job has finished, compare the alignment of the N-terminal visible
\exstep {If you wish,
select and hide a few more columns in the N-terminal region, and submit the alignment to the
service again and explore the effect of local alignment on the non-homologous parts of the
-N-terminal region.}
+N-terminal region.}
+{\bf See the video at:
+\url{http://www.jalview.org/training/Training-Videos}.}
}
+\section{Customising the Parameters used for Alignment}
+
+JABA web services allow you to vary the parameters used when performing a
+bioinformatics analysis. For JABA alignment services, this means you are
+usually able to modify the following types of parameters:
+\begin{list}{$\bullet$}{}
+\item{Amino acid or nucleotide substitution score matrix}
+\item{Gap opening and widening penalties}
+\item{Types of distance metric used to construct guide trees}
+\item{Number of rounds of re-alignment or alignment optimisation}
+\end{list}
+\begin{figure}[htbc]
+\center{
+\includegraphics[width=3in]{images/jvaliwsparamsbox.pdf}
+\caption{{\bf An alignment service's parameter editing dialog box}.}
+\label{jwsparamsdialog} }
+\end{figure}
+
+\subsection{Getting Help on the Parameters for a Service}
+
+Each parameter available for a method usually has a short description, which
+Jalview will display as a tooltip, or as a text pane that can be opened under
+the parameter's controls. In the parameter shown in Figure
+\ref{clustalwparamdetail}, the description was opened by selecting the button on the left hand side.
+
\begin{figure}[htbp]
\begin{center}
\includegraphics[width=2.5in]{images/clustalwparamdetail.pdf}
a preset, then it will be mentioned at the beginning of the job status file shown
in the web service job progress window.
-\subsubsection{Alignment Service Limits}
-Multiple alignment is a computationally intensive calculation. Some JABA server
-services and service presets only allow a certain number of sequences to be
-aligned. The precise number will depend on the server that you are using to
-perform the alignment. Should you try to submit more sequences than a service
-can handle, then an error message will be shown informing you of the maximum
-number allowed by the server.
-
\subsection{User Defined Presets}
Jalview allows you to create your own presets for a particular service. To do
this, select the `{\sl Edit settings and run ...}' option for your service,
description for the parameter set, which will be shown in the tooltip for the
parameter set's entry in the web services menu.
-\begin{figure}[htbc]
-\center{
-\includegraphics[width=3in]{images/jvaliwsparamsbox.pdf}
-\caption{{\bf Jalview's JABA alignment service parameter editing dialog box}.}
-\label{jwsparamsdialog} }
-\end{figure}
-
\subsubsection{Saving Parameter Sets}
When creating a custom parameter set, you will be asked for a file name to save
it. The location of the file is recorded in the Jalview user preferences in the
it will show your custom preset amongst the options available for running the
JABA service.
+%% TODO - reinstate this exercise about reinstating presets
%
% \exercise{Creating and using user defined presets}{\label{createandusepreseex}
% \exstep{Import the file at
% }
% }
+%%TODO just typing a value into parameter slider doesn't actually modify value, return needs to be hit. This is not intuitive (these days) - loss of focus should update values of parameters (e.g. when the run/start job button is hit)
+
\section{Protein Alignment Conservation Analysis}
\label{aacons}
The {\sl Web Service $\Rightarrow$ Conservation} menu controls the computation
-of up to 17 different amino acid conservation measures for the current alignment
+of over 17 different amino acid conservation measures for the current alignment
view. The JABAWS AACon Alignment Conservation Calculation Service, which is used
to calculate these scores, provides a variety of standard measures described by
Valdar in 2002\footnote{Scoring residue conservation. Valdar (2002) {\sl
score developed by Manning et al. in 2008.\footnote{SMERFS Score Manning et al. {\sl BMC
Bioinformatics} 2008, {\bf 9} 51 \href{http://dx.doi.org/10.1186/1471-2105-9-51}{doi:10.1186/1471-2105-9-51}}
-\subsubsection{Enabling and Disabling AACon Calculations}
-When the AACon Calculation entry in the {\sl Web Services $\Rightarrow$
+\subsection{Enabling and Disabling AACon Calculations}
+When the AACon Calculation entry in the {\sl Web Service $\Rightarrow$
Conservation} menu is ticked, AACon calculations will be performed every time
the alignment is modified. Selecting the menu item will enable or disable
automatic recalculation.
-\subsubsection{Configuring which AACon Calculations are Performed}
-The {\sl Web Services $\Rightarrow$ Conservation $\Rightarrow$ Change AACon
+\subsection{Configuring which AACon Calculations are Performed}
+The {\sl Web Service $\Rightarrow$ Conservation $\Rightarrow$ Change AACon
Settings ...} menu entry will open a web services parameter dialog for the
currently configured AACon server. Standard presets are provided for quick and
more expensive conservation calculations, and parameters are also provided to
AACon settings for an alignment are saved in Jalview projects along with the
latest calculation results.
-\subsubsection{Changing the Server used for AACon Calculations}
+\subsection{Changing the Server used for AACon Calculations}
If you are working with alignments too large to analyse with the public JABAWS
server, then you will most likely have already configured additional JABAWS
servers. By default, Jalview will chose the first AACon service available from
-the list of JABAWS servers available. If available, you can switch to use
-another AACon service by selecting it from the {\sl Web Services $\Rightarrow$
-Conservation $\Rightarrow$ Switch Server} submenu.
+the list of JABAWS servers available. You can change the AACon services by
+selecting it from the {\sl Web Service $\Rightarrow$
+Conservation $\Rightarrow$ Change AACon Settings} submenu.
+Alternatively to add new service, go to the desktop window menu and select {\sl Tools $\Rightarrow$
+Preferences $\Rightarrow$ Web Services tab} and add {\sl New Services URL}, then use the {\sl move up} or {\sl move down} buttons
+to reorder the services.
+
\chapter{Analysis of Alignments}
\label{alignanalysis}
Jalview provides support for sequence analysis in two ways. A number of
analytical methods are `built-in', these are accessed from the {\sl Calculate}
alignment window menu. Computationally intensive analyses are run outside
-Jalview {\sl via} web services - these are typically accessed {\sl via} the {\sl
-Web Service} menu, and described in chapter \ref{jvwebservices}.
-In this section, we describe the built-in analysis capabilities common to both
-the Jalview Desktop and the JalviewLite applet.
+Jalview {\sl via} web services - and found under the
+{\sl Web Service} menu. In this section, we describe the built-in analysis
+capabilities common to both the Jalview Desktop and the JalviewJS.
\section{PCA}
-This calculation creates a spatial representation of the similarities within the
-current selection or the whole alignment if no selection has been made. After
-the calculation finishes, a 3D viewer displays the each sequence as a point in
+Principal components analysis calculations create a spatial
+representation of the similarities within the current selection or the whole alignment if no selection has been made. After
+the calculation finishes, a 3D viewer displays each sequence as a point in
3D `similarity space'. Sets of similar sequences tend to lie near each other in
this space.
Note: The calculation is computationally expensive, and may fail for very large
position between each pair of sequences. The basic method is described in the
1995 paper by {\sl G. Casari, C. Sander} and {\sl A. Valencia} \footnote{{\sl
Nature Structural Biology} (1995) {\bf 2}, 171-8.
-PMID: 7749921} and implemented at the SeqSpace server at the EBI.
-
-Jalview provides two different options for the PCA calculation. Protein PCAs are
-by default computed using BLOSUM 62 pairwise substitution scores, and nucleic
-acid alignment PCAs are computed using a score model based on the identity
-matrix that also treats Us and Ts as identical, to support analysis of both RNA
-and DNA alignments. The {\sl Change Parameters} menu also allows the calculation
-method to be toggled between SeqSpace and a variant calculation that is detailed
-in Jalview's built in documentation.\footnote{See
-\url{http://www.jalview.org/help/html/calculations/pca.html}.}
+PMID: 7749921} and implemented at the SeqSpace server at the EBI.\footnote{See \url{http://www.jalview.org/help/html/calculations/pca.html}.}
+%
+% Jalview provides two different options for the PCA calculation: SeqSpace and
+% Jalview mode. In SeqSpace mode, PCAs are computed using the identity matrix, and
+% gaps are treated as 'the unknown residue' (this actually differs from the
+% original SeqSpace paper, and will be adjusted in a future version of Jalview).
+% In Jalview mode, PCAs are computed using the chosen score matrix - which for
+% protein sequences, defaults to BLOSUM 62, and for nucleotides, is the
+% DNA identity matrix that also treats Us and Ts as identical, to support analysis
+% of both RNA and DNA alignments.
\subsubsection{The PCA Viewer}
-PCA analysis can be launched from the {\sl Calculate $\Rightarrow$ Principlal
-Component Analysis} menu option. {\bf PCA requires a selection containing at
-least 4 sequences}. A window opens containing the PCA tool (Figure \ref{PCA}).
+PCA analysis can be launched from the {\sl Calculate $\Rightarrow$ Tree or PCA} menu option.
+{\bf PCA requires a selection containing at
+least 4 sequences}. In the Choose Calculation window, select the {\sl Principal Components Analysis} button and then select {\sl Calculate}
+(Figure \ref{PCA}).
Each sequence is represented by a small square, coloured by the background
colour of the sequence ID label. The axes can be rotated by clicking and
dragging the left mouse button and zoomed using the $\uparrow$ and $\downarrow$
[CTRL]-Click can be used to select multiple sequences.
Labels will be shown for each sequence by toggling the {\sl View $\Rightarrow$
-Show Labels} menu option, and the plot background colour changed {\sl via} the
+Show labels} menu option, and the plot background colour changed {\sl via} the
{\sl View $\Rightarrow$ Background Colour..} dialog box. A graphical
representation of the PCA plot can be exported as an EPS or PNG image {\sl via}
-the {\sl File $\Rightarrow$ Save As $\Rightarrow$ \ldots } submenu.
+the {\sl File $\Rightarrow$ Save as $\Rightarrow$ \ldots } submenu.
\exercise{Principal Component Analysis}
-{ \exstep{Load the alignment at
-\textsf{http://www.jalview.org/tutorial/alignment.fa} }
-\exstep{Select the menu option {\sl Calculate $\Rightarrow$ Principle Component Analysis}.
-A new window will open. Move this window so that the tree, alignment and PCA viewer window are all visible.
+{\label{pcaex}
+\exstep{Load the alignment at
+\textsf{http://www.jalview.org/tutorial/alignment.fa}.}
+\exstep{Select the menu option {\sl Calculate $\Rightarrow$ Tree or PCA..}. in the alignment
+window and a dialogue box will open. Select the Principal Component Analysis option
+and then click the Calculate button.}
+\exstep{Move
+this window within the desktop so that the alignment and PCA viewer windows are visible.
Try rotating the plot by clicking and dragging the mouse on the plot in the PCA window.
-Note that clicking on points in the plot will highlight them on the alignment. }
-\exstep{ Select {\sl Calculate $\Rightarrow$ Calculate Tree $\Rightarrow$
-Neighbour Joining Using BLOSUM62}. A new tree window will appear.
-Click on the tree window. Careful selection of the tree partition location will divide the alignment into a number of groups,
-each of a different colour.
+Note that clicking on points in the plot will highlight the sequences on the
+alignment.}
+\exstep{Use the [ESC] key to deselect sequence selection.
+Select {\sl Calculate $\Rightarrow$ Tree or PCA..}. in the alignment window. In dialogue box select Neighbour
+Joining and in the drop-down list select BLOSUM62. Click the Calculate button
+and a tree window will open.}
+\exstep{Place the mouse cursor on the tree so that the
+tree partition divides the tree into a number of groups, each with a
+different (arbitrarily selected) colour.
Note how the colour of the sequence ID label matches both the colour of
-the partitioned tree and the points in the PCA plot.} }
+the partitioned tree and the points in the PCA plot.}
+{\bf See the video at:
+\url{http://www.jalview.org/training/Training-Videos}.}
+}
\begin{figure}[hbtp]
\begin{center}
\section{Trees}
\label{trees}
+
Jalview can calculate and display trees, providing interactive tree-based
grouping of sequences though a tree viewer. All trees are calculated {\sl via}
-the {\sl Calculate $\Rightarrow$ Calculate Tree $\Rightarrow$ \ldots} submenu.
+the {\sl Calculate $\Rightarrow$ Tree or PCA \ldots} menu option.
Trees can be calculated from distance matrices determined from \% identity or
aggregate BLOSUM 62 score using either {\sl Average Distance} (UPGMA) or {\sl
Neighbour Joining} algorithms. The input data for a tree is either the selected
On calculating a tree, a new window opens (Figure \ref{trees1}) which contains
the tree. Various display settings can be found in the tree window {\sl View}
-menu, including font, scaling and label display options, and the {\sl File
+menu, including font, scaling and label display options. The {\sl File
$\Rightarrow$ Save As} submenu contains options for image and Newick file
export. Newick format is a standard file format for trees which allows them to
be exported to other programs. Jalview can also read in external trees in
{\sl View $\Rightarrow$ Mark Unlinked Leaves} menu option.
+
\begin{figure}
\begin{center}
\includegraphics[width=2.5in]{images/trees1.pdf}
\includegraphics[width=2.5in]{images/trees2.pdf}
\includegraphics[width=1.25in]{images/trees4.pdf}
-\caption{{\bf Calculating Trees} Jalview provides four built in models for calculating trees.
+\caption{{\bf Calculating Trees} Jalview provides a range of options
+for calculating trees.
Jalview can also load precalculated trees in Newick format (right).}
\label{trees1}
\end{center}
\end{figure}
+\begin{figure}
+\begin{center}
+\includegraphics[width=5in]{images/trees3.pdf}
+\caption{{\bf Interactive Trees} The tree level cutoff can be used to designate
+groups in Jalview.}
+\label{trees2}
+\end{center}
+\end{figure}
Clicking on the tree brings up a cursor across the height of the tree. The
sequences are automatically partitioned and coloured (Figure \ref{trees2}). To
and colourschemes on a view, so create a new view ([CTRL-T]) if you wish to
preserve these.
-\begin{figure}
-\begin{center}
-\includegraphics[width=5in]{images/trees3.pdf}
-\caption{{\bf Interactive Trees} The tree level cutoff can be used to designate
-groups in Jalview.}
-\label{trees2}
-\end{center}
-\end{figure}
+
+\exercise{Trees}
+{\label{treeex}
+
+\exstep{Open the alignment at
+\textsf{http://www.jalview.org/tutorial/alignment.fa}.}
+
+\exstep{Select {\sl Calculate $\Rightarrow$ Tree or PCA..}. in the alignment
+window menu and a dialogue box opens. In the tree section select Neighbour
+Joining, in the drop-down list select BLOSUM62 and click the Calculate
+button. A tree window will open.}
+
+\exstep{Click on the
+tree window, a cursor will appear as a vertical line. Note that clicking will
+place this cursor, and divides the tree into a number of groups, each highlighted
+with a different colour. Place the cursor to give about 4 groups.}
+
+\exstep{Place the mouse cursor on a node of the tree to open a tool tip. Double click the node to invert the leaves.
+}
+
+\exstep{In the tree window, select {\sl View $\Rightarrow$ Sort Alignment
+by Tree}. The sequences are reordered to match the order in the tree and groups
+ are formed implicitly. Alternatively in the alignment window, select
+{\sl Calculate $\Rightarrow$ Sort $\Rightarrow$ By Tree Order $\Rightarrow$
+ Neighbour Joining Tree using BLOSUM62 from...}.}
+
+\exstep{Select {\sl Calculate $\Rightarrow$ Tree or PCA..}. in the alignment
+window. In the dialogue box, select Average Distance and in the drop down
+list select BLOSUM62. Click the Calculate button and a new
+tree window will appear. The group colouring makes it easy to see the differences between the two
+trees calculated by the different methods.}
+
+\exstep{With no groups selected in the alignment window, select sequence 2 from
+column 60 to sequence 12 and column 123. Select {\sl Calculate $\Rightarrow$
+Tree or PCA..}. , in the dialogue box select Neighbour Joining and
+BLOSUM62, then click the Calculate button.
+ A tree will appear containing 11 sequences. It has been coloured
+ according to the already selected groups from the first tree and is calculated purely from the residues
+ in the selection.}
+
+Comparing the location of individual sequences between the three trees illustrates the importance of selecting appropriate regions of the
+alignment for the calculation of trees.
+
+{\bf See the video at:
+\url{http://www.jalview.org/training/Training-Videos}.}
+
+}
%\subsubsection{Multiple Views and Input Data recovery from PCA and Tree Viewers}
% move to ch. 3 ?
%Both PCA and Tree viewers are linked analysis windows. This means that their selection and display are linked to a particular alignment, and control and reflect the selection state for a particular view.
-\subsubsection{Recovering input Data for a Tree or PCA Plot Calculation}
+\subsubsection{Recovering input data for a Tree or PCA Plot Calculation}
\parbox[c]{5in}{
The {\sl File $\Rightarrow$ Input Data } option will open a new alignment window containing the original data used to calculate the tree or PCA plot (if available). This function is useful when a tree has been created and then the alignment subsequently changed.
}
\parbox[c]{1.25in}{\centerline{\includegraphics[width=1.25in]{images/pca_fmenu.pdf}
}}
-\subsubsection{Changing the associated View for a Tree or PCA Viewer}
+\subsubsection{Changing the associated view for a Tree or PCA Viewer}
\parbox[c]{4in}{
The {\sl View $\Rightarrow$ Associated Nodes With $\Rightarrow$ .. } submenu is shown when the viewer is associated with an alignment that is involved in multiple views. Selecting a different view does not affect the tree or PCA data, but will change the colouring and display of selected sequences in the display according to the colouring and selection state of the newly associated view.
} \parbox[c]{3in}{\centerline{
better understand the pattern of similarity revealed by the tree and the
variation within the clades partitioned by the grouping. The conservation based
colourschemes and the group associated conservation and consensus annotation
-(enabled using the alignment window's {\sl View $\Rightarrow$ Autocalculated
+(enabled using the alignment window's {\sl Annotations $\Rightarrow$ Autocalculated
Annotation $\Rightarrow$ Group Conservation} and {\sl Group Consensus} options)
can help when working with larger alignments.
-\exercise{Trees}{
-\exstep{Ensure that you have at least 1G memory available in Jalview
-(Either start with this link:
-\href{http://www.jalview.org/services/launchApp?jvm-max-heap=1G}{http://www.jalview.org/services/launchApp?jvm-max-heap=1G},
-or in the Development section of the Jalview web site
-(\href{http://www.jalview.org/development/development-builds}{http://www.jalview.org/development/development-builds})
-in the ``latest official build'' row in the table, go to the
-``Webstart'' column, click on ``G2''.)}
-\exstep{Open the alignment at \textsf{http://www.jalview.org/tutorial/alignment.fa}.
-Select {\sl Calculate $\Rightarrow$ Calculate Tree $\Rightarrow$ Neighbour Joining Using BLOSUM62}. A new tree window will appear.}
-\exstep{Click on the tree window. A cursor will appear. Note that placing this cursor divides the tree into a number of groups by colour.
-Place the cursor to give about 4 groups. In the alignment window, select
-{\sl Calculate $\Rightarrow$ Sort $\Rightarrow$ By Tree Order $\Rightarrow$ Neighbour Joining Tree using BLOSUM62 from ... }. The sequences are
-reordered to match the order in the tree and groups are formed implicitly.
-Alternatively in the tree window, select {\sl View $\Rightarrow$ Sort Alignment
-by Tree}.} \exstep{Select {\sl Calculate $\Rightarrow$ Calculate Tree
-$\Rightarrow$ Neighbour Joining Using \% Identity}. A new tree window will appear. The group colouring makes it easy to see the differences between the two trees, calculated using
- different methods.} \exstep{Select from sequence 2 column 60 to sequence 12 column 123. Select {\sl Calculate $\Rightarrow$ Calculate Tree $\Rightarrow$ Neighbour Joining Using BLOSUM62}. A new tree window will appear. It can be seen that the tree contains 11 sequences. It has been coloured according to the already selected groups from the first tree and is calculated purely from the residues in the selection.
-Comparing the location of individual sequences between the three trees illustrates the importance of selecting appropriate regions of the
-alignment for the calculation of trees. }}
-\exercise{Pad Gaps in Alignment}{
-\exstep{Recover the {\sl Input Data} for the tree you just calculated from the
-{\sl File} menu. In alignment window, ensure that the {\sl Edit $\Rightarrow$
-Pad Gaps } option is {\sl not} ticked, and insert one gap anywhere in the
-alignment.}
-\exstep{Select {\sl Calculate $\Rightarrow$ Calculate Tree $\Rightarrow$
-Neighbour Joining Using BLOSUM62}.
-A warning dialog box {\bf ``Sequences not aligned'' } appears because the sequences input to the tree calculation are of different lengths. }
-\exstep{Select {\sl Edit $\Rightarrow$ tick Pad Gaps } and perform the
-tree calculation again. This time a new tree should appear - because padding
-gaps ensures all the sequences are the same length after editing.
+%\exercise{Pad Gaps in an Alignment}{
+%\exstep{Open the alignment at
+% \textsf{http://www.jalview.org/tutorial/alignment.fa}. In alignment window, ensure that the {\sl Edit $\Rightarrow$ Pad Gaps } option is {\sl not} ticked, and insert one gap anywhere in the
+%alignment.}
+%\exstep{Select {\sl Calculate $\Rightarrow$ Calculate Tree $\Rightarrow$
+%Neighbour Joining Using BLOSUM62}.
-{\sl Pad Gaps } option
-can also be set using
-{\sl Tool $\Rightarrow$ Preference $\Rightarrow$ Editing}. }
+%A warning dialog box {\bf ``Sequences not aligned'' } appears because the
+% sequences input to the tree calculation are of different lengths. }
-}
+%\exstep{Select {\sl Edit $\Rightarrow$ tick Pad Gaps } and perform the
+%tree calculation again. This time a new tree should appear - because padding
+%gaps ensures all the sequences are the same length after editing.}
+%{\sl Pad Gaps } option
+%can be set in Preferences using
+%{\sl Tool $\Rightarrow$ Preference $\Rightarrow$ Editing}.
+
+%{\bf See the video at:
+%\url{http://www.jalview.org/training/Training-Videos}.}
+%}
-\exercise{Tree Based Conservation Analysis}{
+ \exercise{Tree Based Conservation Analysis}{
\label{consanalyexerc}
\exstep{Load the PF03460 PFAM seed alignment using the sequence fetcher.
-Select {\sl Colour $\Rightarrow$ Taylor $\Rightarrow$ By Conservation}
-, and set {\sl Conservation } shading threshold at around 20. }
-\exstep{Build a Neighbour joining tree using Select {\sl Calculate $\Rightarrow$ Calculate Tree $\Rightarrow$
-Neighbour Joining Using BLOSUM62}.}
-\exstep{Use the mouse cursor to select a point on the tree to partition the
-alignment}
-\exstep {{\sl Sort Alignment By Tree} option in the tree viewer submenu to
-re-order the sequences in the alignment using the calculated tree. Examine the
-variation in colouring between different groups of sequences in the alignment
-window.
-
-You may find it easier to browse the alignment if you first uncheck the {\sl
-Annotations $\Rightarrow$ Show Annotations} option, and open the Overview Window
-within the View menu to aid navigation.}
+Select {\sl Colour $\Rightarrow$ Taylor $\Rightarrow$ By Conservation}, set
+ {\sl Conservation} shading threshold at around 20. }
+ \exstep{Build a Neighbour joining tree by selecting {\sl Calculate
+ $\Rightarrow$ Tree or PCA..}. in the alignment window. In the dialogue box, select Neighbour
+Joining and in the drop-down
+list select BLOSUM62, then click the Calculate button.}
+\exstep{Use the cursor to select a point on the tree to partition the
+alignment into groups.}
+\exstep {Select {\sl View $\Rightarrow$ Sort Alignment By Tree} option in the
+tree window to re-order the sequences in the alignment.
+Examine the variation in colouring between different groups of sequences in the
+ alignment window. }
+\exstep {You may find it easier to browse the alignment if you first uncheck
+ the {\sl Annotations $\Rightarrow$ Show Annotations} option. Open the
+Overview Window from the View menu to aid navigation.}
\exstep{Try changing the colourscheme of the residues in the alignment to
BLOSUM62 (whilst ensuring that {\sl Apply Colour to All Groups} is selected).}
-{\sl Note: You may want to save the alignment and tree as a project file, since it is used in the next few exercises. } }
+{\sl Note: You may want to save the alignment and tree as a project file, since
+it is used in the next set of exercises. }
+
+{\bf See the video at:
+\url{http://www.jalview.org/training/Training-Videos}.}
+}
\subsection{Redundancy Removal}
-The redundancy removal dialog box is opened using the {\sl Edit $\Rightarrow$ Remove Redundancy\ldots} option in the alignment menu. As its menu option placement suggests, this is actually an alignment editing function, but it is convenient to describe it here. The redundancy removal dialog box presents a percentage identity slider which sets the redundancy threshold. Aligned sequences which exhibit a percentage identity greater than the current threshold are highlighted in black. The [Remove] button can then be used to delete these sequences from the alignment as an edit operation\footnote{Which can usually be undone. A future version of Jalview may allow redundant sequences to be hidden, or represented by a chosen sequence, rather than deleted.}.
+The redundancy removal dialog box is opened using the {\sl Edit $\Rightarrow$ Remove Redundancy\ldots} option
+in the alignment menu. As its menu option placement suggests, this is actually an alignment editing function,
+but it is convenient to describe it here. The redundancy removal dialog box presents a percentage identity
+slider which sets the redundancy threshold. Aligned sequences which exhibit a percentage identity greater
+than the current threshold are highlighted in black. The [Remove] button can then be used to delete these
+sequences from the alignment as an edit operation.
\begin{figure}
\begin{center}
\includegraphics[width=5.5in]{images/redundancy.pdf}
\end{center}
\label{removeredundancydialog}
-\caption{The Redundancy Removal dialog box opened from the edit menu. Sequences that exceed the current percentage identity threshold and are to be removed are highlighted in black.}
+\caption{The Redundancy Removal dialog box opened from the edit menu. Sequences that exceed the current percentage identity
+threshold and are to be removed are highlighted in black.}
\end{figure}
\exercise{Remove Redundant Sequences}{
-
-\exstep{Re-use or recreate the alignment and tree which you worked with in the
-tree based conservation analysis exercise (exercise \ref{consanalyexerc}). In
-the alignment window, you may need to deselect groups using Esc key.}
+\label{redundantex}
+\exstep{Using the alignment generated in the previous exercise (exercise
+\ref{consanalyexerc}).
+In the alignment window, you may need to deselect groups using Esc key.}
\exstep{In the {\sl Edit} menu select {\sl Remove Redundancy} to open the
Redundancy threshold selection dialog. Adjust the redundancy threshold value, start
at 50 and increase the value to 65. Sequences selected will change colour in the Sequence ID panel. Select ``Remove'' to
remove the sequences that are more than 65\% similar under this alignment.}
-\exstep{Select the Tree viewer's {\sl View $\Rightarrow$ Mark Unlinked Leaves} option, and note that the removed sequences are now prefixed with a * in the tree view.}
-\exstep{Use the [Undo] button in the Redundancy threshold selection dialog box
+\exstep{From the tree window, select {\sl View $\Rightarrow$
+Mark Unlinked Leaves} option, and note that the removed sequences are now prefixed with a * in the tree view.}
+ \exstep{Use the [Undo] button in the Redundancy threshold selection dialog box
to recover the sequences. Note that the * symbols disappear from the tree display.}
-\exstep{Experiment with the redundancy removal and observe the relationship between the percentage identity threshold and the pattern of unlinked nodes in the tree display.}
+\exstep{Experiment with the redundancy removal and observe the relationship between the percentage identity threshold and
+the pattern of unlinked nodes in the tree display.}
}
\exercise{Group Conservation Analysis}{
+\label{conservationex}
\exstep{Re-use or recreate the alignment and tree which you worked with in the
-tree based conservation analysis exercise (exercise \ref{consanalyexerc})}
-\exstep{In the {\sl View} menu, select {\sl New View} to create a new
-view. Ensure the annotation panel is displayed ({\sl Show annotation} in {\sl Annotations} menu). Enable the
+tree based conservation analysis exercise (exercise \ref{consanalyexerc}).}
+\exstep{In the {\sl View} menu in the alignment window, select {\sl New View} to
+create a new view. Ensure the annotation panel is displayed ({\sl Annotations $\Rightarrow$ Show Annotations} is toggled on). Enable the
display of {\sl Group Consensus} option by checking {\sl Group Consensus} in the {\sl Annotation $\Rightarrow$
-Autocalculated Annotation } submenu in the sequence alignment window.}
+Autocalculated Annotation $\Rightarrow$
+Group Consensus} submenu.}
+
\exstep{Displaying the sequence
logos will make it easier to see the different residue populations within each
-group. Activate the logo by right clicking on the Consensus annotation row to
-open the context menu and select the {\sl Show Logo} option.}
-\exstep{In the column alignment ruler, select a column exhibiting about 50\%
-conservation that lies within the central conserved region of the alignment.
-(Column 74 is used in the Tree video).}
+group. Activate the logo by right clicking the name of the Consensus annotation to
+open the context menu and select the {\sl Show Logo} option. Alter its size, by moving the cursor onto the annotation row, right clicking the
+mouse and dragging it up. Alter its position, by right clicking the name of the Consensus annotation
+and dragging it up to the top of the annotations.}
+\exstep{In the column alignment ruler, select a column exhibiting with about 50\%
+ of its residues conserved ({\em ie. about 50\% in the consensus histogram})
+ that lies within the central conserved region of the alignment.
+(Column 74 is used in \href{https://youtu.be/m-PjynicXRg}{the Tree video}).}
\exstep{Subdivide the alignment
-according to this selection using {\sl Select $\Rightarrow$ Make groups for selection}.}
+according to this selection using {\sl Select $\Rightarrow$ Make Groups for Selection}.}
\exstep{Re-order the alignment according to the new groups that have been
defined by selecting {\sl Calculate $\Rightarrow$ Sort $\Rightarrow$
By Group}.
-
Click on the group annotation row IDs to select groups exhibiting a
specific mutation.}
\exstep{Select another column exhibiting about 50\% conservation
overall, and subdivide the alignment further. Note that the new groups
inherit the names of the original groups, allowing you to identify the
-combination of mutations that resulted in the subdivision.
-}
+combination of mutations that resulted in the subdivision.}
\exstep{Clear the groups, and try to subdivide the alignment using two
non-adjacent columns.
both of the columns that you wish to use to subdivide the alignment.}}
\exstep{Switch back to the original view, and experiment with subdividing
the tree groups made in the previous exercise.}
+{\bf See the video at:
+\url{http://www.jalview.org/training/Training-Videos}.}
}
\begin{figure}[]
\end{figure}
-\chapter{Working with 3D structures}
-\label{3Dstructure}
-Jalview can interactively view 3D structure using Jmol or Chimera. Setting in
-the Structure window within Preferences determine whether Jmol or Chimera is
-the default choice of structure viewer.
-
% To review, Chapter \ref{featannot} describes the mechanisms provided by
% Jalview for interactive creation of sequence and alignment annotation, and how
% they can be displayed, imported and exported and used to reorder the alignment. Chapter
% and Section \ref{workingwithrna} covers the visualization,
% editing and analysis of RNA secondary structure.
-\section{Working with Structures}
+\chapter{Working with 3D structures}
+\label{3Dstructure}
\label{wkwithstructure}
-Jalview facilitates the use of protein structures for the analysis of alignments
-by providing a linked view of structures associated with sequences in
-the alignment. The Java based molecular viewing program Jmol\footnote{See the
-Jmol homepage \url{http://www.jmol.org} for more information.} has been
-incorporated\footnote{Earlier versions of Jalview included MCView - a simple
-main chain structure viewer. Structures are visualized as an alpha carbon trace
-and can be viewed, rotated and coloured in a structure viewer and the results
-interpreted on a sequence alignment.} which enables sophisticated molecular
-visualizations to be prepared and investigated alongside an analysis of
-associated sequences.
-PDB format files can be imported directly or structures can be retrieved from
-the European Protein Databank (PDBe) using the Sequence Fetcher (see
-\ref{fetchseq}).
-
-\subsection{Automatic Association of PDB Structures with Sequences}
-Jalview can automatically determine which structures are associated with a
-sequence in a number of ways.
-\subsubsection{Discovery of PDB IDs from Sequence Database Cross-references}
-If a sequence has an ID from a public database that contains cross-references to
-the PDB, such as Uniprot. Right-click on any sequence ID and select {\sl Structure $\Rightarrow$
-Associate Structure with Sequence $\Rightarrow$ Discover PDB IDs } from the context menu (Figure \ref{auto}). Jalview will attempt to associate the
-sequence with a Uniprot sequence and from there discover any associated PDB
-structures. This takes a few seconds and applies to all sequences in the
-alignment which have valid Uniprot IDs. On moving the cursor over the sequence
-ID the tool tip\footnote{Tip: The sequence ID tooltip can often become large for
-heavily cross referenced sequence IDs. Use the {\sl View $\Rightarrow$ Sequence
-ID Tooltip $\Rightarrow$ } submenu to disable the display of database cross
-references or non-positional features. } now shows the Uniprot ID and any
-associated PDB structures.
+Jalview facilitates the use of 3D structure data for the analysis of alignments
+by providing a linked view of structures associated with the aligned sequences.
+It also allows sequence, secondary structure and B-factor data to be imported
+from structure files, and supports the use of the EMBL-EBI's SIFTS database to
+construct accurate mappings between UniProt protein sequences and structures
+retrieved from the PDB.
+
+\section{Molecular graphics systems supported by Jalview}
+Jalview can interactively view 3D structure using Jmol, a Java based molecular
+viewing program\footnote{See the Jmol homepage \url{http://www.jmol.org} for
+more information.} integrated with Jalview.\footnote{Earlier
+versions of Jalview included MCView - a simple main chain structure viewer.
+Structures are visualized as an alpha carbon trace and can be viewed, rotated
+and coloured using the sequence alignment.} It also supports the use of UCSF
+Chimera, a powerful molecular graphics system that needs separate installation.
+Jalview can also read PDB and mmCIF format files directly to extract sequences
+and secondary structure information, and retrieve records from the European
+Protein Databank (PDBe) using the Sequence Fetcher (see \ref{fetchseq}).
+
+\subsection{Configuring the default structure viewer}
+\label{configuring3dviewer}
+To configure which viewer is used when creating a new
+structure view, open the Structures preferences window {\sl via} {\sl Tools $\Rightarrow$ Preferences\ldots} and
+select either JMOL or CHIMERA as the default viewer. If you select Chimera,
+Jalview will search for the installed program, and if it cannot be found,
+you will be prompted to locate the Chimera binary, or alternately, open the UCSF
+Chimera download page to obtain the software.
+
+\section{Automatic Association of PDB Structures with Sequences}
+Jalview will attempt to automatically determine which structures are associated
+with a sequence via its ID, and any associated database references. To do this
+for a particular sequence or the current selection, open the Sequence ID popup
+menu and select {\sl View 3D Structure}, to open the 3D Structure Chooser.
+%(Figure\ref{auto}).
+
+When the structure chooser is first opened, if no database identifiers are
+available, Jalview will automatically perform a database reference
+retrieval (See \ref{fetchdbrefs}) to discover identifiers for the
+sequences to use to search the PDB. This can take a
+few seconds for each sequence and will be performed for all selected
+sequences.\footnote{After this is done, you can see the added database
+references in a tool tip by mousing over the sequence ID. You can use the {\sl
+View $\Rightarrow$ Sequence ID Tooltip $\Rightarrow$ Show Db References }
+submenu option to enable or disable these data in the tooltip.}
+
+Once the retrieval has finished, the structure chooser dialog will show any
+available PDB entries for the selected sequences.
-\begin{figure}[htbp]
-\begin{center}
-%TODO fix formatting
-\parbox{1.5in}{
-{\centering
-\begin{center}
-\includegraphics[width=1.5in]{images/auto1.pdf}
-\end{center}}
-} \parbox{3.25in}{
-{\centering
-\begin{center}
-\includegraphics[scale=0.5]{images/auto2.pdf}
-\end{center}
-}
-} \parbox{1.5in}{
-{\centering
-\begin{center}
-\includegraphics[width=1.5in]{images/auto3.pdf}
-\end{center}
-}
-}
-
-\caption{{\bf Automatic PDB ID discovery.} The tooltip (left) indicates that no PDB structure has been associated with the sequence.
-After PDB ID discovery (center) the tool tip now indicates the Uniprot ID and
-any associated PDB structures (right).}
-\label{auto}
-\end{center}
-\end{figure}
+%
+% \begin{figure}[htbp]
+% \begin{center}
+% %TODO fix formatting
+% \begin{center}
+% \includegraphics[width=3.5in]{images/pdbstructurechooser.pdf}
+% \end{center}
+%
+%
+% \caption{{\bf The PDB Structure Chooser dialog.} }
+% \label{auto}
+% \end{center}
+% \end{figure}
-\subsubsection{Drag-and-Drop Association of PDB Files with Sequences by Filename
+\subsection{Drag-and-Drop Association of PDB Files with Sequences by Filename
Match}
\label{multipdbfileassoc}
-If one or more PDB files stored on your computer are dragged from their location
-on the file browser onto an alignment window, Jalview will search the alignment
-for sequences with IDs that match any of the files dropped onto the alignment.
-If it discovers matches, a dialog like the one in Figure
-\ref{multipdbfileassocfig} is shown, giving the option of creating associations
-for the matches.
+If you have PDB files stored on your computer {\bf named the same way as the
+sequences in the alignment}, then you can drag them from their location on the
+file browser onto an alignment window. Jalview will search the alignment for
+sequences with IDs that match any of the files, and offer a dialog like the one
+in Figure \ref{multipdbfileassocfig}.
If no associations are made, then sequences extracted
from the structure will be simply added to the alignment. However, if only
sequence within a local directory. Check out
\href{http://issues.jalview.org/browse/JAL-801}{Jalview issue 801}}
+After associating sequences with PDB files, you can view the PDB structures by
+opening the Sequence ID popup
+menu and selecting {\sl View 3D Structure}. The PDB files you loaded will be
+shown in the {\bf Cached Structures} view, after selecting it from the drop down
+menu in the dialog box.
+
% there is no mention of the other footnote (#3) that appears saying: Tip: The sequence ID tooltip can often become large for heavily cross-referenced sequence IDs. Use the ...
% JBP: yes there is - under 'Discovery of ' subsection.
\begin{figure}[htbp]
\end{figure}
-\subsection{Viewing Structures}
-\label{viewAllStructures}
-The structure viewer can be launched in two ways from the sequence ID context
-menu. To view a particular structure associated with a sequence in the
-alignment, simply select it from popup menu's associated structures submenu in
-{\sl Structure $\Rightarrow$ View Structure $\Rightarrow$ $<$PDB ID$>$}. The
-second way is most useful if you want to view all structural data available for
-a set of sequences in an alignment. If any of the {\bf currently selected}
-sequences have structures associated, the {\sl Structure } submenu of the
-sequence ID popup menu will include an option to {\sl View {\bf N}
-structures}. Selecting this option will open a new structure view containing
-the associated structures superposed according to the alignment.
-
-In both cases, each structure to be displayed will be downloaded or loaded from
+\section{Viewing Structures}
+\label{structurechooser}
+The structure viewer is launched via the Sequence ID context
+menu. To view structures associated with a sequence or a selected set of
+sequences in the alignment, simply right click the mouse to open the context
+menu, and select {\sl 3D Structure data \ldots} to open the Structure Chooser
+dialog box.
+
+If any of
+the {\bf currently selected} sequences have structures in the PDB,
+they will appear in the Structure Chooser dialog box. The structures can be ranked by
+different parameters, but are by default ordered according to their PDB
+quality score.
+
+To view one or more structures, simply click {\sl
+View} to open a structure viewer containing the structures selected in the
+dialog. If several structures were picked, these will be shown
+superposed according to the alignment.
+You may find Jalview has already picked the best structure - using one of the
+criteria shown in the dropdown menu (e.g. 'Best Quality', which is picked by
+default). However, you are free to select your own.
+
+The structure(s) to be displayed will be downloaded or loaded from
the local file system, and shown as a ribbon diagram coloured according to the
-associated sequence in the current alignment view (Figure \ref{structure}
-(right)). The structure can be rotated by clicking and dragging in the structure
+associated sequence in the current alignment view (Figure \ref{structure}). The structure can be rotated by clicking and dragging in the structure
window. The structure can be zoomed using the mouse scroll wheel or by
[SHIFT]-dragging the structure.
+
Moving the mouse cursor over a sequence to which the structure is linked in the
alignment view highlights the respective residue's sidechain atoms. The
sidechain highlight may be obscured by other parts of the molecule. Similarly,
\begin{figure}[htbp]
\begin{center}
-\parbox{3in}{
+\parbox{4in}{
{\centering
\begin{center}
-\includegraphics[scale=0.5]{images/structure1.pdf}
+\includegraphics[width=4in]{images/structure1.pdf}
\end{center}
}
}
-\parbox{3.2in}{
+\parbox{2.2in}{
{\centering
\begin{center}
-\includegraphics[width=3in]{images/structure2.pdf}
+\includegraphics[width=2.2in]{images/structure2.pdf}
\end{center}
}
}
-\caption{{\bf Structure visualization} The structure viewer is launched from the sequence ID context menu (left) and allows the structure to be visualized using the embedded Jmol molecular viewer (right). }
+\caption{{\bf Structure visualization} Structure viewers are launched from
+the 3D Structure chooser dialog (left). Jalview shows the displayed structures
+coloured according the alignment view (right). }
\label{structure}
\end{center}
\end{figure}
in the structure viewer can be saved as an EPS or PNG with the {\sl File
$\Rightarrow$ Save As $\Rightarrow$ \ldots} submenu, which also allows the raw
data to be saved as PDB format. The mapping between the structure and the
-sequence (How well and which parts of the structure relate to the sequence) can
+sequence (how well and which parts of the structure relate to the sequence) can
be viewed with the {\sl File $\Rightarrow$ View Mapping} menu option.
\subsubsection{Using the Jmol Visualization Interface }
automatic application of colour schemes when new structure data is added, or
when associated alignment views are modified.
-\exercise{Viewing Structures in Jmol viewer}{\label{viewingstructex}
+%%TODO: no mapping when using N&W alg.
+
+\exercise{Viewing Structures with the integrated Jmol Viewer}{
+\label{viewingstructex}
\exstep{Load the alignment at
-\textsf{http://www.jalview.org/examples/exampleFile.jar}.}
+\textsf{http://www.jalview.org/examples/exampleFile.jvp}.}
\exstep{Right-click on the
sequence ID label of {\sl FER1\_SPIOL} to open
-the context menu. Select {\sl 3D Structure}, this
-opens a Structure Chooser window, select { \sl 1A70} and click { ok}.
-
-{\sl Note: the Structure Chooser interface
-provides a smart technique for selecting PDB structures by queryingthe meta-data
-of structures. Extra information can be including in this window by checking boxes
-in the ``Configure Displayed Columns'' tab}.
+the ID popup menu and select {\sl 3D Structure Data}. After a short pause, a
+Structure Chooser dialog will open for the sequence, listing available
+structure data from the PDB. Select { \sl 1a70} from the list and click {\sl
+New View}.
+
+{\sl Note:} The Structure Chooser dialog presents available PDB structures
+by querying the EMBL-EBI's PDBe web API. Extra information can be
+including in this window by checking boxes in the columns of the {\sl Customise Displayed Options} tab.
% JBP Note: Bug JAL-1238 needs to be fixed ASAP
}
\exstep{By default the Jmol
structure viewer opens in the Jalview desktop. Rotate the molecule by clicking
and dragging in the structure viewing box.
Zoom with the mouse scroll wheel. }
-\exstep{Roll the
-mouse cursor along the {\sl FER1\_SPIOL} sequence in the alignment.
-Note that if a residue in the sequence maps to one in the structure, a label
-will appear next to that residue in the structure viewer.}
-\exstep{Move the mouse over the structure. In the Jmol viewer, placing the mouse over a
+\exstep{In the Jmol viewer, placing the mouse over a
part of the structure will bring up a tool tip indicating the name and number of that residue.
In the alignment window, the corresponding residue in the sequence is
highlighted in black.}
+\exstep{Roll the
+mouse cursor along the {\sl FER1\_SPIOL} sequence in the alignment window.
+If a residue in the sequence maps to one in the structure, the residue molecular shape is visible in the structure viewer.}
\exstep{Clicking the alpha carbon toggles the highlight and residue label on and
off.
Try this by clicking on a set of three or four adjacent residues so that the labels are persistent, then finding where they are in the sequence. }
\exstep{In the structure viewer menu, select {\sl Colours $\Rightarrow$ Background
Colour\ldots} and choose a suitable colour.
-Press OK to apply this.}
+Press {\sl OK} to apply this.}
\exstep{Select {\sl File $\Rightarrow$ Save As $\Rightarrow$ PNG} and save the
image. On your computer, view this with a suitable program. }
-\exstep{Select
-{\sl File $\Rightarrow$ View Mapping} from the structure viewer menu.
+\exstep{Select {\sl File $\Rightarrow$ View Mapping} from the structure viewer menu.
A new window opens showing the residue by residue alignment between the sequence and the structure.}
-\exstep{Select {\sl File $\Rightarrow$ Save $\Rightarrow$ PDB file} and choose a new filename to save the PDB file.
-Once the file is saved, open the location in your file browser (or explorer window) and drag the PDB file that you just saved on
-to the Jalview desktop (or load it from the {\sl Jalview Desktop $\Rightarrow$ Input Alignment $\Rightarrow$ From File } menu).
-Verify that you can open and view the associated structure from the sequence ID
-pop-up menu's {\sl 3D Structure } submenu in the new alignment window.}
-
-\exstep{In the Jmol window, right click on the structure window and explore the
-menu options. Try to change the style of molecular display - for example by
-using the {\sl Jmol $\Rightarrow$ Select (n) $\Rightarrow$ All} command (where {\sl n} is the number of residues selected), and then the {\sl Jmol $\Rightarrow$ Style $\Rightarrow$ Scheme $\Rightarrow$ Ball and Stick} command.}
-\exstep{In the alignment window, use the {\sl File $\Rightarrow$ Save As .. }
-function to save the alignment as a Jalview Project. Now close the alignment and the structure view, and load the project file you just saved.
-Verify that the Jmol display is as it was when you just saved the file.}}
-
-\exercise{Setting Chimera as the default 3D Structure Viewer}{\label{viewingchimera}
+\exstep{In the structure window, select {\sl File $\Rightarrow$ Save as $\Rightarrow$ PDB file} and enter a filename to save the PDB file.
+Once the file is saved, open the location in your file browser and drag the PDB file that you just saved onto the Jalview desktop.
+(Or load it from the Jalview Desktop menu using
+{\sl File $\Rightarrow$ Input Alignment $\Rightarrow$ From File}).
+Verify that you can open and view the associated structure from new alignment window using the sequence ID
+context menu's {\sl 3D Structure } submenu (as step {\sl b)}.}
+
+\exstep{In the Jmol window, right click on the background to open the
+Jmol menu options. Explore this, for example by
+{\sl Select (n) $\Rightarrow$ All} command (where {\sl n} is the number of residues selected),
+and then the {\sl Style $\Rightarrow$ Scheme $\Rightarrow$ Ball and Stick} command.}
+\exstep{In the alignment window, use the {\sl File $\Rightarrow$ Save as.. }
+function to save the alignment as a Jalview Project (jvp). Now close the alignment and the structure view, and load the project file you just saved.
+Verify that the Jmol display is as it was when you just saved the file.}
+{\bf See the video at:
+\url{http://www.jalview.org/training/Training-Videos}.}
+}
+
+\exercise{Aligning Structures using the Ferredoxin Sequence Alignment}{
+\label{superpositionex}
+
+\exstep{Continue with the Jalview project created in exercise
+\ref{viewingstructex}}
+
+\exstep{Select the FER1\_MAIZE sequence (near bottom of the alignment). Right-click the ID label to open the context menu,
+select {\sl $\Rightarrow$ 3D Structure Data}.}
+
+\exstep{This opens the Structure Chooser dialog, pick 1gaq from the list.
+Make sure the {\sl Superpose} option is checked before clicking the {\bf Add}
+button. This superimpose the structure associated with
+FER1\_MAIZE with the one associated with FER1\_SPIOL.
+
+{\sl Note:} The Jmol view should update to show both structures, and one will be
+moved on to the other. If this doesn't happen, use the Align function in the
+Jmol submenu.
+}
+
+\exstep{Create a new view on the alignment ({\sl View $\Rightarrow$ New View}), and hide all but columns 121
+through to 132 ({\sl View $\Rightarrow$ Hide $\Rightarrow$
+All but selected region}).}
+\exstep{Select the newly created view in the structure window using {\sl Jmol $\Rightarrow$ Superpose
+With } submenu, and then recompute the superposition with {\sl Jmol
+$\Rightarrow$ Superpose Structures}.
+
+{\sl Note:} how the molecules shift position when superposed with only a small
+region of the alignment.}
+
+\exstep{Compare RMSDs obtained when superimposing molecules with
+columns 121-132 and with the whole alignment. The RMSD report can be
+viewed by right clicking the mouse on Jmol window, and select {\sl
+Console} from the menu (if nothing is shown, recompute the superposition after
+displaying the console).}
+\exstep{Which view do you think give the best 3D superposition, and why ?} }
+
+\exercise{Setting Chimera as the default 3D Structure Viewer}{
+\label{viewingchimera}
Jalview supports molecular structure
visualization using both Jmol and Chimera 3D viewers. Jmol is the default
viewer, however Chimera can be set up as the default choice from Preferences.
\exstep{Close the Jalview program, from the
{\sl Desktop menu} select {\sl Jalview $\Rightarrow$ Quit Jalview}. Then reopen
Jalview, Chimera should open as the default viewer.}
-{\s Note: The Jmol structure viewer sits within the Jalview desktop; however the Chimera structure viewer
-sits outside the Jalview desktop and a Chimera
-view window sits inside the Jalview desktop.} }
+If this does not work then you may need to set the {\sl Path to Chimera program} in {\sl Preferences}.
+{\sl Note:} The Jmol structure viewer sits within the Jalview desktop. However
+the Chimera structure viewer sits outside the Jalview desktop and a Chimera
+view window sits inside the Jalview desktop.
+
+{\bf See the video at: \url{http://www.jalview.org/training/Training-Videos}.}}
+
\subsection{Superimposing Structures}
\label{superposestructs}
biochemical properties of a given molecule are significantly different to its
homologues. When structure data is available, comparing the shapes of molecules
by superimposing them enables substructure that may impart different behaviour
-to be quickly identified. The identification of optimal 3D superpositions
+to be quickly identified. The identification of optimal 3D superposition
involves aligning 3D data rather than sequence symbols, but the result can
still be represented as a sequence alignment, where columns indicate positions
in each molecule that should be superposed to recreate the optimal 3D alignment.
Jalview can employ Jmol's 3D fitting routines\footnote{See
\href{http://chemapps.stolaf.edu/jmol/docs/?ver=12.2$\#$compare}{http://chemapps.stolaf.edu/jmol/docs/?ver=12.2$\#$compare}
for more information.} to recreate 3D structure superpositions based on the
-correspondences defined by one or more sequence alignments involving structures shown in the Jmol display. Superposition based on the currently displayed alignment view happens automatically if a
-structure is added to an existing Jmol display using the {\sl Structure
-$\Rightarrow$ View PDB Structure $\Rightarrow$ ..}. A new Jmol view containing
-superposed structures can also be created using the {\sl Structure
-$\Rightarrow$ View all {\bf N} PDB Structures} option (when {\bf {\sl N}}
-$>$ 1) if the current selection contains two or more sequences with associated
+correspondences defined by one or more sequence alignments involving structures shown in the Jmol display.
+Superposition based on the currently displayed alignment view
+ happens automatically if a
+structure is added to an existing Jmol display using
+the {\sl 3D Structure } option in the Sequence ID popup menu to open the
+Structure Chooser dialog box.
+Select the structures required and select {\sl View}. A new Jmol view
+opens containing superposed structures if the current selection contains two or more sequences with associated
structures.
\subsubsection{Obtaining the RMSD for a Superposition}
the cartoon style, with other parts of the molecule drawn in wireframe. The Jmol
console, which has been opened after the superposition was performed, shows the
RMSD report for the superposition.
-Full information about the superposition is also outputed to the Jalview
+Full information about the superposition is also reported on the Jalview
console.\footnote{The Jalview Java Console is opened from {\sl Tools
$\Rightarrow$ Java Console} option in the Desktop's menu bar} This output also
includes the precise atom pairs used to superpose structures.
directly compared.
In order to recompute a superposition after changing a view or editing the
-alignment, select the {\sl Jmol $\Rightarrow$ Align sequences } menu option. The {\sl
-Jmol $\Rightarrow$ Superpose with ..} submenu allows you to choose which of the
+alignment, select the {\sl Jmol $\Rightarrow$ Align Structures} menu option.
+The {\sl Jmol $\Rightarrow$ Superpose with ..} submenu allows you to choose which of the
associated alignments and views are to be used to create the set of
correspondences. This menu is useful when composing complex superpositions
involving multi-domain and multi-chain complexes, when correspondences may be
Note that these menu options appear when you have two or more structures in one Jmol viewer.
-\begin{figure}[htbp]
-\begin{center}
-\includegraphics[width=5.5in]{images/fdxsuperposition.pdf}
-\caption{{\bf Superposition of two ferredoxin structures.} The alignment on the
-left was used by Jalview to superpose structures associated with the
-FER1\_SPIOL and FER1\_MAIZE sequences in the alignment. Parts of each structure
-used for superposition are rendered as a cartoon, the remainder rendered in
-wireframe. The RMSD between corresponding positions in the structures before and
-after the superposition is shown in the Jmol console.}
-\label{mstrucsuperposition}
-\end{center}
-\end{figure}
-
\subsection{Colouring Structure Data Associated with Multiple Alignments and
Views} Normally, the original view from which a particular structure view was
opened will be the one used to colour structure data. If alignments involving
Note that the {\sl Select many views} option is useful if you have different
views that colour different areas or domains of the alignment. This option is
-further explored in Section \ref{complexstructurecolours}.
+further explored in exercise \ref{complexstructurecolours}.
+
+\begin{figure}[htbp]
+\begin{center}
+\includegraphics[width=5.5in]{images/fdxsuperposition.pdf}
+\caption{{\bf Superposition of two ferredoxin structures.} The alignment on the
+left was used by Jalview to superpose structures associated with the
+FER1\_SPIOL and FER1\_MAIZE sequences in the alignment. Parts of each structure
+used for superposition are rendered as a cartoon, the remainder rendered in
+wireframe. The RMSD between corresponding positions in the structures before and
+after the superposition is shown in the Jmol console.}
+\label{mstrucsuperposition}
+\end{center}
+\end{figure}
\begin{figure}[htbp]
\begin{center}
\end{center}
\end{figure}
-\exercise{Aligning Structures using the Ferredoxin
-Sequence Alignment}{\label{superpositionex}
-
-\exstep{Continue with the Jalview project created in exercise
-\ref{viewingstructex}. Use the {\sl Discover PDB IDs} function to retrieve PDB
-IDs associated with the FER1\_MAIZE sequence.}
-\exstep{Once discovery has completed, use the {\sl
-View PDB Structure} submenu to view one of the PDB file associated with
-FER1\_MAIZE (eg. 3B2F)
-Jalview will give you the option of aligning the structure to the one already
-open. To superimpose the structure associated with FER1\_MAIZE with the one
-associated with FER1\_SPIOL, press the {\bf Yes} button.
-
-{\sl The Jmol view will update to show both structures, and one will be
-moved on to the other. If this doesn't happen, use the Align function in the Jmol submenu}}
-\exstep{Create a new view on the alignment, and hide all but columns 121
-through to 132.}
-\exstep{Use the {\sl Jmol} submenu to
-recompute the superposition using just columns 121-132 of the alignment
-(The easiest way to achieve this is to select column 121-132 and in the View
-menu selected ``All but selected region'' from the Hide options).
-
-{\sl Note how the molecules shift position when superposed using a short part of
-the two structures.}}
-\exstep{Compare the initial and final RMSDs for superimposing molecules with
-the small section and with the whole alignment. (The RMSD report can be
-viewed by right clicking the mouse on Jmol window, and select ``Show" and ``Measurements") Which view do you think give the best 3D
-superposition, and why ?} }
-
\subsubsection{Colouring Complexes}
\label{complexstructurecolours}
The ability to control which multiple alignment view is used to colour
\exstep{Download the PDB file at
\textsf{\url{http://www.jalview.org/tutorial/DNMT1\_MOUSE.pdb}} to your desktop.
-
This is the biological unit for PDB ID 3pt6, as identified by the PDBe's PISA
server.}
-\exstep{Launch the Jalview desktop and ensure you have at least 256MB of
-free memory available.
-{\sl Use the following webstart link:
-\href{http://www.jalview.org/services/launchApp?jvm-max-heap=1G}{http://www.jalview.org/services/launchApp?jvm-max-heap=1G}.}
-{\sl Alternatively in the Development section of the Jalview web site
-(\href{http://www.jalview.org/development/development-builds}{http://www.jalview.org/development/development-builds})
-in the ``latest official build'' row in the table, go to the
-``Webstart'' column, click on ``G2''.}}
-\exstep{Retrieve the following {\bf full} PFAM alignments: PF02008, PF01426
-(make sure you select the {\sl PFAM {\bf (Full)}} source). These will each be retrieved into their own alignment window.} \exstep{Drag the URL or file of the structure you downloaded in
-step 1 onto one of the alignments to associate it with the mouse sequence in
-that Pfam domain family.}
-\exstep{For every DNMT1\_MOUSE sequence in the alignment, use the sequence
-ID popup menu's {\sl Structure} submenu to view the DNMT1\_MOUSE structure for the associated mouse sequence. When given the option, {\bf view all of the structures in the same Jmol viewer}. Check the contents of the {\sl View $\Rightarrow$ Colour by ..} submenu to see what alignments can be used to
-colour the sequence.}
-\exstep{Repeat the previous two steps for each of
-the other alignments. In each case, when performing the `View DNMT1\_MOUSE.pdb'
-step, Jalview will ask if you wish to create a new Jmol view. You should
-respond `No', {\bf ensuring that each sequence fragment is associated with the same Jmol view}.}
-\exstep{Pick a different colourscheme for each alignment, and use the {\sl
-Colour by ..} submenu to ensure they are all used to colour the complex shown
-in the Jmol window.}
+
+\exstep{Retrieve the following PFAM alignments from the {\bf PFAM (full)} source
+:
+
+PF02008; PF01426; PF00145 (retrieve each into their own alignment window).}
+
+\exstep{Drag the URL or file of the structure you
+downloaded in step 1 onto one of the alignments to associate it with the mouse sequence in that Pfam domain family.}
+
+\exstep{Locate every DNMT1\_MOUSE sequence in the
+alignment by opening the Find dialog box via {\sl Select
+$\Rightarrow$ Find}. Search using the text DNMT1\_MOUSE. Open the Structure Chooser dialog box
+by right clicking the mouse on
+sequence name to open the context menu and select {\sl
+$\Rightarrow$ 3D Structure Data}.
+Select `Cached Structures' from
+the drop-down menu in the Structure Chooser dialog box and select the
+DNMT1\_MOUSE.pdb structure, and click {\sl New View}.
+
+{\em Part of the newly opened structure will be coloured the same way as
+the associated DNMT1\_MOUSE sequence is in the alignment view.}
+
+{\bf WARNING: do not select all sequences and open the Structure Chooser
+!} {\em This will cause Jalview to attempt to discover all structures for
+sequences in the alignment.}
+}
+\exstep{Repeat the previous two steps for the other two
+ alignments. For those, after selecting the DNMT1\_MOUSE.pdb structure you
+ should select {\bf `Add'} to ensure each domain alignment is associated
+ with the {\bf same} Jmol view. }
+
+\exstep{Pick a different
+colourscheme in each alignment window for each alignment using the {\sl Colour $\Rightarrow$ Colour by...} submenu to
+ensure each of the complexes shown in the Jmol window are coloured.
+{\sl The different shading schemes will highlight regions of strong
+physicochemical conservation on corresponding domains in the structure.}
+}
+
\exstep{The final step needed to reproduce the shading in Figure
\ref{mviewalcomplex} is to use the {\sl Colour $\Rightarrow$ By
-Annotation } option in each alignment window to shade the alignment by the
-{\bf Conservation} annotation row. This function was described in section
-\ref{colourbyannotation}.
+Annotation\ldots } option in each alignment window to shade the alignment by the
+{\bf Conservation} annotation row (introduced in section
+\ref{colourbyannotation}).
-Ensure that you first disable the {\sl View $\Rightarrow$ Show Features} menu option, or you may not see any colour changes in the associated structure.
+{\sl Note:} Ensure that you first disable the {\sl View $\Rightarrow$ Show Features} menu
+option, or you may not see any colour changes in the associated structure.
-{\sl Note: Choose a different shading scheme for each
-alignment so that the regions of strong physicochemical conservation are highlighted. This
-kind of shading will reveal conserved regions of interaction between domains
-in the structure.}}
-\exstep{Save your work as a Jalview project and verify that it can be opened again by starting another Jalview Desktop instance, and dragging the saved project into the desktop window.}
+{\sl Examine the regions strongly coloured at the interfaces between each
+protein domain, and the DNA binding region. What do you think these patterns
+mean? } }
+\exstep{Save your work as a Jalview project and verify that it can be opened
+again by starting another Jalview Desktop instance, and dragging the saved
+project into the Desktop window.}
% {\sl Note: This exercise relies on new features introduced in Jalview 2.7. If
% you notice any strange behaviour when trying out this exercise, it may be a
}
% TODO
-\chapter{Protein Prediction Analysis}
+\chapter{Protein sequence analysis and structure prediction}
\label{proteinprediction}
+
+Many of Jalview's sequence feature and annotation capabilities were developed to
+allow the results of sequence based protein structure prediction methods to be
+visualised and explored. This chapter introduces services integrated with the
+Jalview Desktop for predicting protein secondary structure and protein disorder.
+
\section{Protein Secondary Structure Prediction}
\label{protsspredservices}
Protein secondary structure prediction is performed using the
-Jpred\footnote{{\sl ``The Jpred 3 Secondary Structure Prediction Server''} Cole, C., Barber, J. D. and Barton, G. J. (2008) {\sl Nucleic Acids Research} {\bf 36}, (Web Server Issue) W197-W201
+Jpred\footnote{{\sl ``The Jpred 3 Secondary Structure Prediction Server''} Cole,
+C., Barber, J. D. and Barton, G. J. (2008) {\sl Nucleic Acids Research} {\bf
+36}, (Web Server Issue) W197-W201
{\sl ``Jpred: A Consensus Secondary Structure Prediction Server''} Cuff, J. A.,
Clamp, M. E., Siddiqui, A. S., Finlay, M. and Barton, G. J. (1998) {\sl
the same criteria as above, then the alignment will be used for a JPred
prediction on the first sequence in the set (that is, the one that appears first in the alignment window).
\end{list}
-Jpred is launched in the same way as the other web services. Select {\sl Web
-Services $\Rightarrow$ Secondary Structure Prediction $\Rightarrow$ JNet
-Secondary Structure Prediction}\footnote{JNet is the Neural Network based
-secondary structure prediction method that the JPred server uses.} from the
-alignment window menu (Figure \ref{jpred}).
-A status window opens to inform you of the progress of the job. Upon completion, a new alignment window opens and the Jpred
-predictions are included as annotations. Consult the Jpred documentation for
-information on interpreting these results.
+
+\exercise{Secondary Structure Prediction}{
+\label{secstrpredex}
+
+{\sl Note:} The annotation panel can get quite busy during this exercise. Try
+hiding some annotations rows by right clicking
+the mouse in the annotation label panel and select the ``Hide this row'' option.
+The Annotations dropdown menu on the alignment window also provides options for
+reordering and hiding autocalculated and sequence associated annotation.
+
+\exstep{ Open the alignment at \url{http://www.jalview.org/tutorial/alignment.fa}. Select the sequence {\sl FER\_MESCR} by
+clicking on the sequence ID. Then select {\sl Web Service $\Rightarrow$
+Secondary Structure Prediction $\Rightarrow$ JPred Secondary Structure
+Prediction} from the alignment window menu. A status window will appear and after some time (about 2-4 min)
+a new window with the JPred prediction will appear. The results
+from the prediction are visible in the annotation panel. JPred secondary
+structure prediction annotations are examples of sequence-associated alignment annotation. {\sl Note:} The number of sequences in the results
+window is many more than in the original alignment as
+JPred performs a PSI-BLAST search to expand the prediction dataset.}
+% TODO: check how long this takes - about 2 mins once it gets on the cluster.
+\exstep{
+Select a different sequence and perform a JPred prediction in the same way.
+There will probably be minor differences in the predictions.
+}
+\exstep{
+Select the sequence used in the second sequence prediction by clicking on its
+name in the sequence ID panel, and copy {\sl ([CTRL] or [CMD]-C)} and then
+paste it {\sl [CTRL] or [CMD]-V)} into the first prediction window. You
+can now compare the two predictions as the annotations associated with the
+sequence has also been copied across.
+% which is described in Section \ref{seqassocannot} below.
+}
+\exstep{
+Select and hide some columns in one of the alignment profiles that were returned
+from the JNet service, and then submit the profile for prediction again.
+}
+\exstep{
+When you get the result, verify that the prediction has not been made for the
+hidden parts of the profile by clicking the mouse on column ruler and right click to open the
+context menu and select {\sl Reveal All}. The JPred reliability scores
+differ from the prediction made on the full profile.
+}
+\exstep{
+In the original alignment that you loaded in step {\sl a}, select {\bf all}
+sequences ([CTRL]-A or [CMD]-A), then open the {\sl Sequence ID $\Rightarrow$ Selection $\Rightarrow$ Add
+Reference Annotation} submenu
+by right clicking the mouse to open the context menu.
+{\bf All} the JPred predictions for the sequences will now be visible in the
+original alignment window.}
+ {\bf Homework:} Go back to the last step of exercise \ref{annotatingalignex} and
+follow the instructions to view the Jalview annotations file created from the annotations
+generated by the JPred server for your sequence.
+\bf See the video at:
+\url{http://www.jalview.org/training/Training-Videos}.}
\begin{figure}[htbp]
\begin{center}
\includegraphics[width=2.25in]{images/jpred1.pdf}
\includegraphics[width=3in]{images/jpred2.pdf}
-\caption{{\bf Secondary Structure Prediction} Status (left) and results (right) windows for JNet predictions. }
+\caption{{\bf Secondary Structure Prediction} Status (left) and results (right)
+windows for JPred predictions. }
\label{jpred}
\end{center}
\end{figure}
-\subsubsection{Hidden Columns and JNet Predictions}
+
+Jpred is launched in the same way as the other web services. Select {\sl Web
+Service $\Rightarrow$ Secondary Structure Prediction $\Rightarrow$ JPred
+Secondary Structure Prediction}\footnote{JNet is the Neural Network based
+secondary structure prediction method that the JPred server uses.} from the
+alignment window menu (Figure \ref{jpred}).
+A status window opens to inform you of the progress of the job. Upon completion, a new alignment window opens and the Jpred
+predictions are included as annotations. Consult the Jpred documentation for
+information on interpreting these results.
+
+\subsection{Hidden Columns and JPred Predictions}
\label{hcoljnet}
Hidden columns can be used to exclude parts of a sequence or profile from the
input sent to the JNet service. For instance, if a sequence is known to include
a large loop insertion, hiding that section prior to submitting the JNet
-prediction can produce different results. In some cases, these secondary structure predictions can be more reliable for sequence on either side of the insertion\footnote{This, of course, cannot be guaranteed.}. Prediction results returned from the service will
-be mapped back onto the visible parts of the sequence, to ensure a single frame
-of reference is maintained in your analysis.
+prediction can produce different results. In some cases, these secondary
+structure predictions can be more reliable for sequence on either side of the
+insertion\footnote{This, of course, cannot be guaranteed.}. Prediction results
+returned from the service will be mapped back onto the visible parts of the
+sequence, to ensure a single frame of reference is maintained in your analysis.
\section{Protein Disorder Prediction}
\label{protdisorderpred}
Disordered regions in proteins were classically thought to correspond to
``linkers'' between distinct protein domains, but disorder can also play a role in
-function. The {\sl Web Services $\Rightarrow$ Disorder} menu in the alignment window
+function. The {\sl Web Service $\Rightarrow$ Disorder} menu in the alignment window
allows access to protein disorder prediction services provided by the configured
JABAWS servers.
+\begin{figure}[htbp]
+\begin{center}
+\includegraphics[width=5in]{images/disorderpredannot.pdf}
+\caption{{\bf Annotation rows for several disorder predictions on a sequence}. A
+zoomed out view of a prediction for a single sequence. The sequence is shaded to highlight disordered regions (brown and grey), and the line plots below the Sequence show the raw scores for various disorder predictors. Horizontal lines on each graph mark the level at which disorder predictions become significant. }
+\label{alignmentdisorderannot}
+\end{center}
+\end{figure}
+
\subsection{Disorder Prediction Results}
Each service operates on sequences in the alignment to identify regions likely
to be unstructured or flexible, or alternately, fold to form globular domains.
thresholding (described in Section \ref{featureschemes}) can be used to
highlight differences in disorder prediction across aligned sequences.
-\exercise{Secondary Structure Prediction}{
-\label{secstrpredex}
-\exstep{ Open the alignment at \url{http://www.jalview.org/tutorial/alignment.fa}. Select the sequence {\sl FER\_MESCR} by
-clicking on the sequence ID. Then select {\sl Web Services $\Rightarrow$ Secondary Structure Prediction $\Rightarrow$ JNet Secondary Structure Prediction}
-from the alignment window menu. A status window will appear and after some time (about 2-4 min) a new window with the JPred prediction will appear.
-Note that the number of sequences in the results window is many more than in the original alignment as
-JNet performs a PSI-BLAST search to expand the prediction dataset. The results
-from the prediction are visible in the annotation panel. Jnet secondary
-structure prediction annotations are examples of sequence-associated alignment annotation. }
-% TODO: check how long this takes - about 2 mins once it gets on the cluster.
-\exstep{
-Select a different sequence and perform a JNet prediction in the same way. There will probably be minor differences in the predictions.
-}
-\exstep{
-Select the sequence used in the second sequence prediction by clicking on its
-name in the sequence ID panel, and copy {\sl (CTRL or Command + C)} and then
-paste it {\sl (CTRL or Command + V)} into the first prediction window. You
-can now compare the two predictions as the annotations associated with the
-sequence has also been copied across.
-% which is described in Section \ref{seqassocannot} below.
-}
-\exstep{
-Select and hide some columns in one of the alignment profiles that were returned
-from the JNet service, and then submit the profile for prediction again.
-}
-\exstep{
-When you get the result, verify that the prediction has not been made for the
-hidden parts of the profile (by clicking the mouse on column ruler and right click to open the
-context menu and select {\sl Reveal All}), and that the JPred reliability scores
-differ from the prediction made on the full profile.
-}
-\exstep{
-In the original alignment that you loaded in step 1, select {\bf all}
-sequences, then open the {\sl Sequence ID $\Rightarrow$ Selection } submenu
-by right clicking the mouse to open the context menu, and select the {\sl Add
-Reference Annotation} option.
-
-{\bf All} the JNet predictions for the sequences will now be visible in the
-original alignment window.
-
-{\sl Note: The annotation panel can get quite busy and it may be
-helpful to hide some of the annotations rows, by right clicking the mouse in
-the annotation label panel and select ``Hide this row'' option in the context
-menu}.} }
-
-\begin{figure}[htbp]
-\begin{center}
-\includegraphics[width=5in]{images/disorderpred.pdf}
-\caption{{\bf Shading alignment by sequence disorder}. Alignment of Interleukin IV homologs coloured with Blosum62 with protein disorder prediction sequence features overlaid, shaded according to their score. Borderline disordered regions appear white, reliable predictions are either Green or Brown depending on the type of disorder prediction. }
-\label{alignmentdisorder}
-\end{center}
-\end{figure}
-\subsubsection{Navigating Large Sets of Disorder Predictions}
+\subsection{Navigating Large Sets of Disorder Predictions}
Figure \ref{alignmentdisorderannot} shows a single sequence annotated with
a range of disorder predictions. Disorder prediction annotation rows are
the associated sequence in the alignment display, and double clicking will
select that sequence.
-\begin{figure}[htbp]
-\begin{center}
-\includegraphics[width=5in]{images/disorderpredannot.pdf}
-\caption{{\bf Annotation rows for several disorder predictions on a sequence}. A
-zoomed out view of a prediction for a single sequence. The sequence is shaded to highlight disordered regions (brown and grey), and the line plots below the Sequence show the raw scores for various disorder predictors. Horizontal lines on each graph mark the level at which disorder predictions become significant. }
-\label{alignmentdisorderannot}
-\end{center}
-\end{figure}
-
-
\subsection{Disorder Predictors provided by JABAWS 2.0}
For full details of each predictor and the results that Jalview can display,
please consult
\textbf{REMARK465} ``Missing
coordinates in X-ray structure as defined by remark465 entries in PDB.
Nonassigned electron densities most often reflect intrinsic disorder,
-and have been used early on in disorder prediction.'' Features give
+and have been used early on in disorder prediction'' . Features give
range(s) of residues predicted as disordered, and annotation rows gives
raw value for each residue. Values over 0.1204 indicates disorder.
+\begin{figure}[htbp]
+\begin{center}
+\includegraphics[width=5in]{images/disorderpred.pdf}
+\caption{{\bf Shading alignment by sequence disorder}. Alignment of Interleukin IV homologs coloured with Blosum62 with protein
+disorder prediction sequence features overlaid, shaded according to their score. Borderline disordered regions appear white,
+reliable predictions are either Green or Brown depending on the type of disorder prediction. }
+\label{alignmentdisorder}
+\end{center}
+\end{figure}
+
\subsubsection{RONN {\sl a.k.a.} Regional Order Neural Network}
\href{http://www.strubi.ox.ac.uk/RONN}{RONN} employs an approach
known as the `bio-basis' method to predict regions of disorder in
values are structured.
{\bf Disordered region} sequence features are created marking mark range(s) of residues with positive first order derivatives, and
-\textbf{Globular Domain} features mark long stretches of order. \textbf{Dydx} annotation rows gives the first order derivative of smoothed score. Values above 0 indicates
+\textbf{Globular Domain} features mark long stretches of order. \textbf{Dydx} annotation rows give the first order derivative of smoothed score. Values above 0 indicates
residue is disordered.
\textbf{Smoothed Score and Raw Score} annotation rows give the smoothed and raw scores used to create the differential signal that
by default, but can be shown by right-clicking on the alignment
annotation panel and selecting \textbf{Show hidden annotation}.
-\exercise{Protein Disorder Prediction}{
-%\label{protdispredex}
-
-\exstep{Open the alignment at
-\url{http://www.jalview.org/tutorial/interleukin7.fa}. } also available at
-
-
-\url{http://www.jalview.org/tutorial/training-materials/2014/Dundee/Oct/interleukin7.fa}
-
-\exstep{Run the DisEMBL disorder predictor {\slvia} the {\slWeb Services
-$\Rightarrow$ Disorder Prediction } submenu.}
+\exercise{Protein Disorder Prediction}
+{
+\label{protdisorderex}
+{\sl Note:} Before starting this exercise, make sure you enable the \protect{{\sl Add
+Temperature Factor annotation to alignment}} option in your Structures preferences
+({\sl Tool $\Rightarrow$ Preferences $\Rightarrow$ Structure)}.
-\exstep{Use {\sl Sequence ID $\Rightarrow$ Structure $\Rightarrow$ Discover PDB
-IDs} to retrieve all the PDB structures for the sequences.}
+\exstep{Open the alignment from
+\url{http://www.jalview.org/tutorial/interleukin7.fa}. }
-\exstep{Open and align
-the structures for all sequences.
+\exstep{Run the DisEMBL disorder predictor {\sl via} the {\sl Web Service
+$\Rightarrow$ Disorder Prediction $\Rightarrow$ Disembl with defaults}.}
-{\sl Hint: see \ref{viewAllStructures} to see how to do this.}}
+\exstep{Select all the sequences ([CTRL]-A or [CMD]-A). Open the Structure Chooser by placing
+the mouse in the Sequence ID panel, right clicking the mouse and select
+{\sl$\Rightarrow$ 3D Structure Data\ldots }. Select all structures in the list.
+Hit the {\sl New View} button to retrieve and show all PDB structures for the sequences.}
\exstep{Compare the disorder predictions to the structure data by mapping any
available temperature factors to the alignment {\sl via} the {\sl Sequence ID
Popup $\Rightarrow$ Selection $\Rightarrow$ Add reference annotation} option.}
-\exstep{Apply the IUPred disorder prediction method}
-\exstep{Use the {\sl Per
-sequence option} in the {\sl Colour $\Rightarrow$ By annotation \ldots} dialog to shade
-the sequences by the long and short disorder predictors.
-Do the two methods agree with the structure ?}}
+\exstep{Features on sequences can conceal other colouring. This can be
+toggled off by selecting {\sl View
+$\Rightarrow$ Show Sequence Features} in the alignment window menu.}
+\exstep{Apply the IUPred disorder prediction method. Tick the
+{\sl Per sequence option} in the {\sl Colour $\Rightarrow$ By annotation \ldots} dialog
+box. Then shade the sequences by the long and short disorder predictors. {\sl
+Note} how well the disordered regions predicted by each method agree
+with the structure.}
+\bf See the video at:
+\url{http://www.jalview.org/training/Training-Videos}.}
\chapter{DNA and RNA Sequences}
\label{dnarna}
and alignments. Jalview recognises nucleotide sequences and alignments based on
the presence of nucleotide symbols [ACGT] in greater than 85\% of the
sequences. Built in codon-translation tables can be used to translate ORFs
-into peptides for further analysis. EMBL nucleotide records retrieved {\sl via} the
+into peptides for further analysis. ENA nucleotide records retrieved {\sl via} the
sequence fetcher (see Section \ref{fetchseq}) are also parsed in order to
identify codon regions and extract peptide products. Furthermore, Jalview
records mappings between protein sequences that are derived from regions of a
\subsection{Linked DNA and Protein Views}
\parbox{3.5in}{
-Views of alignments involving DNA sequences are linked to views of alignments containing their peptide products in a similar way to views of protein sequences and views of their associated structures. Peptides translated from cDNA that have been fetched from EMBL records for DNA contigs are linked to their `parent' coding regions. Mousing over a region of the peptide highlights codons in views showing the original coding region.
+Views of alignments involving DNA sequences are linked to views of alignments containing their peptide products in a similar way to views of protein sequences and views of their associated structures. Peptides translated from cDNA that have been fetched from ENA records for DNA contigs are linked to their `parent' coding regions. Mousing over a region of the peptide highlights codons in views showing the original coding region.
}\parbox{3in}{
\begin{center}
%\begin{figure}[htbp]
}
-\subsection{Coding Regions from EMBL Records}
+\subsection{Coding Regions from ENA Records}
-Many EMBL records that can be retrieved with the sequence fetcher contain exons.
-Coding regions will be marked as features on the EMBL nucleotide sequence, and
+Many ENA records that can be retrieved with the sequence fetcher contain exons.
+Coding regions will be marked as features on the ENA nucleotide sequence, and
Uniprot database cross references will be listed in the tooltip displayed when
the mouse hovers over the sequence ID. Uniprot database cross references
-extracted from EMBL records are sequence cross references, and associate a
+extracted from ENA records are sequence cross references, and associate a
Uniprot sequence's coordinate system with the coding regions annotated on the
-EMBL sequence. Jalview utilises cross-reference information in two ways.
+ENA sequence. Jalview utilises cross-reference information in two ways.
\subsubsection{Retrieval of Protein or DNA Cross References}
-The {\sl Calculate $\Rightarrow$ Get Cross References } function is only available when Jalview recognises that there are protein/DNA cross-references present on sequences in the alignment. When selected, it retrieves the cross references from the alignment's dataset (a set of sequence and annotation metadata shared between alignments) or using the sequence database fetcher. This function can be used for EMBL sequences containing coding regions to open the Uniprot protein products in a new alignment window. The new alignment window that is opened to show the protein products will also allow dynamic highlighting of codon positions in the EMBL record for each residue in the protein product(s).
+The {\sl Calculate $\Rightarrow$ Get Cross References } function is only available when Jalview recognises that there are protein/DNA cross-references present on sequences in the alignment. When selected, it retrieves the cross references from the alignment's dataset (a set of sequence and annotation metadata shared between alignments) or using the sequence database fetcher. This function can be used for ENA sequences containing coding regions to open the Uniprot protein products in a new alignment window. The new alignment window that is opened to show the protein products will also allow dynamic highlighting of codon positions in the ENA record for each residue in the protein product(s).
-\subsubsection{Retrieval of Protein DAS Features on Coding Regions}
+\subsubsection{Retrieval of Protein Features on Coding Regions}
-The Uniprot cross-references derived from EMBL records can be used by Jalview to visualize protein sequence features directly on nucleotide alignments. This is because the database cross references include the sequence coordinate mapping information to correspond regions on the protein sequence with that of the nucleotide contig. Jalview will use the Uniprot accession numbers associated with the sequence to retrieve features, and then map them onto the nucleotide sequence's coordinate system using the coding region location.
+The Uniprot cross-references derived from ENA records can be used by Jalview to visualize protein sequence features directly on nucleotide alignments.
+This is because the database cross references include the sequence coordinate mapping information to correspond regions on the protein sequence with that of the nucleotide contig.
+Jalview will use the Uniprot accession numbers associated with the sequence to retrieve features, and then map them onto the nucleotide sequence's coordinate system using
+the coding region location.
\begin{figure}[htbp]
\begin{center}
\label{dnadasfeatures}
\includegraphics[width=5in]{images/dnadasfeatures.pdf}
-\caption{Uniprot and PDB sum features retrieved {\sl via} DAS and mapped onto
-coding regions of EMBL record V00488 (an earlier version of Jalview is shown
+\caption{Uniprot and PDB sum features retrieved and mapped onto
+coding regions of ENA record V00488 (an earlier version of Jalview is shown
here).}
\end{center}
\exercise{Visualizing Protein Features on Coding Regions}
{
-\exstep{Use the sequence fetcher to retrieve EMBL record D49489.}
+\label{protfeatureex}
+\exstep{Use the sequence fetcher to retrieve ENA record D49489.}
\exstep{Ensure that {\sl View $\Rightarrow$ Show Sequence Features} is checked and change the
-alignment view format to Wrapped mode so the distinct exons can be seen.}
+alignment view format to {\sl Wrapped} mode so the distinct exons can be seen.}
\exstep{Open the {\sl DAS Settings} tab in the {\sl Sequence Feature Settings\ldots}
window {\sl View $\Rightarrow$ Features setting}
and fetch features for D49489 from the Uniprot reference server,
\subsection{Performing RNA Secondary Structure Predictions}
Secondary structure consensus calculations can be performed by enabling the
-VIENNA service {\sl via} the {\sl Web Services $\Rightarrow$ Secondary
+VIENNA service {\sl via} the {\sl Web Service $\Rightarrow$ Secondary
Structure} menu. These consensus structures are created by analysing the
covariation patterns in all visible sequences on the alignment. For more
information see the VIENNA documentation.
\includegraphics[width=5in]{images/rnaViennaServiceWindow.pdf}
\caption{Secondary structure consensus calculations can be performed by enabling the
-VIENNA service {\sl via} the {\sl Web Services $\Rightarrow$ Secondary
+VIENNA service {\sl via} the {\sl Web Service $\Rightarrow$ Secondary
Structure} menu.}
\end{center}
\exercise{Viewing RNA Structures}
{ \label{viewingrnaex}
-\exstep{Import RF00162 from Rfam (Full) using Fetch sequence(s) option in File
-menu.}
-\exstep{Select {\sl Colour by RNA Helices} to shade the alignment by
-the secondary structure annotation provided by Rfam.}
+\exstep{Import RF00162 from the Rfam (Seed) source using {\sl Fetch sequence(s)}
+from the Desktop's File menu.}
+
+\exstep{Select {\sl Colour by RNA Helices} to
+shade the alignment by the secondary structure annotation provided by Rfam.}
\exstep{Open VARNA with {\sl Structure $\Rightarrow$ View Structure
$\Rightarrow$ RNA Secondary Structure}.
In the VARNA Structures Manager toggle between (i) secondary structure
(alignment) (with gaps) and (ii) trimmed secondary structure (alignment).
Explore the difference between trimmed and untrimmed views.
-Click on different residues and located them in the sequence alignment window.}
+Click on different residues in the VARNA diagram - you should also see them
+highlighted and selected in the sequence alignment window.}
\exstep{In the VARNA Structures Manager, right click on display window to
-bring up the pop up context menu. Investigate option within File, Export, Display
-and Edit sections.}
+bring up the pop up context menu. Explore the options within the File, Export,
+Display and Edit sections.
+
+{\em VARNA views are stored in Jalview project files, in the same way as 3D
+structure views produced by Jmol and Chimera.}}
-\exstep{Perform a secondary structure prediction in {\sl Web Services}. Enable
-the VIENNA consensus calculation via the {\sl Web Services} menu and select
-Change RNAAIiFold settings option.}
+\exstep{Enable the calculation and display of an RNAAliFold secondary structure
+prediction for the alignment by selecting {\sl Web Service $\Rightarrow$ Secondary
+Structure Prediction $\Rightarrow$ RNAAliFold }.}
% Compare this with the annotationline provided by Rfam.
-\exstep{Edit the VIENNA calculation settings to show
+\exstep{Edit the RNAAliFold calculation settings to show
Base Pair probabilities. Explore how editing the alignment affects the consensus
calculation.}
-\exstep{Import 2GIS from PDB database using Fetch sequence(s) option.}
+\exstep{Import 2GIS from the PDB database into a new window with {\sl Fetch
+sequence(s)}.}
+
\exstep{Click on a sequence in Sequence ID panel and select {\sl Structure
$\Rightarrow$ View Structure $\Rightarrow$ 2GIS}, to view the structure in Jmol
window. Click on different residues and located them in the sequence alignment window.}
+
%\exstep{Add and link a Jmol structure view
%for Bacillus\_amyloliquef.9 for 3NPB (from the PDB). Display the secondary
%structure along-side the consensus structure for the alignment by adding
}
\chapter{Webservices}
+
+\section{What are Web Services ?}
+
\label{jvwebservices}
The term ``Webservices'' refers to a variety of data exchange
mechanisms based on HTTP.\footnote{HTTP: Hyper-Text Transfer Protocol.}
which were introduced in in Section \ref{fetchseq}, provide sequence and
alignment data. They can also be used to add sequence IDs to an alignment
imported from a local file, prior to further annotation retrieval, as described
-in Section \ref{featuresfromdb}. A second type of one way service is provided
-by Jalview's DAS sequence feature retrieval system, which is described
-in Section \ref{dasfretrieval}.
+in Section \ref{featuresfromdb}.
% The final type of one way service are sequence
% and ID submission services.
% exemplified by the `Envision2 Services' provided
progress of running jobs.
-\subsection{JABA Web Services for Sequence Alignment and Analysis}
+\section{JABA Web Services for Sequence Alignment and Analysis}
\label{jabaservices}
JABA stands for ``JAva Bioinformatics Analysis'', which is a system developed
by Peter Troshin and Geoff Barton at the University of Dundee for running
\subsection{Running your own JABA Server}
You can download and run JABA on your own machine using the `VMWare' or
-VirtualBox virtual machine environments. If you would like to learn how to do
+VirtualBox virtual machine environments. If you would like to do
this, there are full instructions at the
\href{http://www.compbio.dundee.ac.uk/jabaws/}{JABA web site}.
WARNING: This is large (about 300MB) and will take some time to download.
}
\exstep{Unpack the archive's contents to a place on your machine with at least
-2GB of free space.
-
-(On Windows, right click on the archive, and use the 'Extract archive..' option).
+2GB of free space (On Windows, right click on the archive, and use the 'Extract
+archive..' option).
}
\exstep{Open the newly extracted directory and double click the VMWare virtual
machine configuration file (jabaws.vcf). This will launch the VMWare player.
git://source.jalview.org / jalview-manual.git / blobdiff