\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 2.10.1}
+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
+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 1.8
+Manual Version 1.9.2
% post CLS lifesci course on 15th January
% draft. Remaining items are AACon, RNA visualization/editing and Protein disorder analysis exercises.
-15th February 2017
+
+26th April 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.
alignments for web sites such as Pfam.\footnote{\url{http://pfam.xfam.org}}
-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. It also
+\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
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 Chimera structure displays,
+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.
\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 concerns the
+appropriate, typically at the end of each section. 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 and opening Chimera, building
-and viewing trees and PCA plots, and using trees for sequence conservation analysis. An overview of
+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. 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}
respectively.
-Section \ref{featannot} details the creation and visualization of sequence and alignment annotation. 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.
+Section \ref{featannot} details the creation and visualization of sequence,
+features and alignment annotation. 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.
%^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 JalviewLite applet, 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 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
\exstep {If
you are having trouble, it may help changing the browser you are using, as the browsers and
its version may affect this process.}
-\exstep{To deactivate the opening of the 4 demo sequences during the launch, go
+\exstep{To disable opening of the demonstration project during the launch, go
to the {\sl Tools $\Rightarrow$
Preferences...} menu on the desktop. A `Preference'
dialog box opens, untick the box adjacent to the `Open file' entry in the
\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 (a downward arrow on the right hand side of the dialog
-box) to view the files, select exampleFile\_2\_7.jar, then click {\sl OK}.}
-{\bf Note:} Should you want to load your own
-sequence during the launch process, then go
+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.
-
-
-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 the jnlp 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 See the video at:
\url{http://www.jalview.org/Help/Getting-Started}.}
\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 key}
- [Fn]-F2.
+ is used to switch between these two modes.
+
+ {\em Note:} On MacBooks and other laptops with compact keyboards, you may need
+ to press the {\bf function key [Fn]} when pressing any of the numbered function
+ keys. So to toggle between keyboard and normal mode, press [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}
%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).
{\bf Note:} To view Jalview's comprehensive on-line help documentations select
{\sl Help} in desktop menu, clicking on {\sl 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.
+Documentation window. Select topics from the navigation panel on the left hand
+side and use the Search tab to locate specific key words.
{\sl\bf See the video at:
\url{http://www.jalview.org/Help/Getting-Started}.}
%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
+ 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})
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.
+
+To begin retrieving data, select {\sl File $\Rightarrow$ Fetch Sequence(s) \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}
\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
+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
\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.
-
+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.}
+
\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 save the file to your desktop.
+your web browser and save the 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 {\sl OK} to load.}
-\exstep{{\bf Loading sequences by `Drag and Drop' / `Cut and Paste':}
+$\Rightarrow$ Input Alignment $\Rightarrow$ From File} from the desktop menu.
+Select the file and click {\sl OK} to load.}
-(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.
+\exstep{{\bf Loading sequences by `Drag and Drop' / `Cut and Paste':}
-(ii) Test the differences
-between (a) dragging the sequence onto the Jalview desktop and (b)
-dragging the sequence onto an existing alignment window.
+(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.
+}
-(iii) 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.)}
+\exstep{{\bf The text editor:}
-\exstep{{\bf The text editor:} (i) 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 the {\sl Paste to New Window} menu
-option.
+(i) Open the alignment.fa file using text editor.
+Copy the sequence text into the clipboard using [CTRL]-A and then [CTRL]-C.
-(ii) 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).
+(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.
-(iii) 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.}
+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 Sequence(s)...} from the Desktop. The {\sl Select Database
-Retrieval Source} dialog will open showing all the database sources. Select the
-{\bf PFAM seed} database and click {\sl OK}.
+$\Rightarrow$ Fetch Sequence(s)...} 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}.
-(ii)Once a source has been selected, the {\sl New
+(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.}
-Several database IDs can be loaded by using semicolons to separate them.}
+$\Rightarrow$ Overview Window.}}
{\bf See the video at:
\url{http://www.jalview.org/Help/Getting-Started}} }
%[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}
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{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.}
\exstep{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 See the video at: \url{http://www.jalview.org/Help/Getting-Started}.}
-}
+$\Rightarrow$ 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.
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,
-but adds to the column selection.
+it adds to the column selection.
Selected columns are indicated by red highlighting in the ruler bar (Figure \ref{selectcols}).
%[fig 13]
\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
+sequence ID panel. The same techniques can be used as for columns above ([SHIFT]-Click for continuous and [CTRL]-Click {\sl (Or
+[CMD]-Click for Mac)}) to select discontinuous
ranges of sequences (Figure \ref{selectrows}).
%[fig 14]
\exercise{Making Selections and Groups}{
\label{exselect}
-\exstep{Close windows.
-
-Load the ferredoxin alignment ({\bf PF03460} from {\bf PFAM (seed)}).
+\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
\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 exported and saved by right clicking the
-mouse when on the text area to open the Sequence ID context 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 that opens, try manually editing the
alignment before clicking the {\sl New Window} button. This opens the
\end{figure}
\exercise{Reordering the Alignment}{
+\label{reorderex}
\exstep{Close windows.
Load the ferredoxin alignment ({\bf PF03460} from {\bf PFAM (seed)}).
\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}
\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.
+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.
\exercise{Hiding and Revealing Regions}{
-\exstep{Close windows.
-
-Load the ferredoxin alignment ({\bf PF03460} from {\bf PFAM (seed)}).
+\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
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).
-\subsection{Sliding Sequences}
-Pressing the [$\leftarrow$] or [$\rightarrow$] arrow keys when one or more
-sequences are selected will ``slide'' the entire selected sequences to the left
-or right (respectively). Slides occur regardless of the region selection -
-which, for example, allows you to easily reposition misaligned subfamilies
-within a larger alignment.
-% % 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.
-
\newpage
\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}.
+ \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. }
backwards and replay the edits you have made.}
}
+\subsection{Sliding Sequences}
+Pressing the [$\leftarrow$] or [$\rightarrow$] arrow keys when one or more
+sequences are selected will ``slide'' the entire selected sequences to the left
+or right (respectively). Slides occur regardless of the region selection -
+which, for example, allows you to easily reposition misaligned subfamilies
+within a larger alignment.
+% % 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{Keyboard Edits}
-{This continues on from the previous exercise, 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 Window users:} Please {\em only use} [SHIFT]-[SPACE] in this
\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}
} \parbox[c]{3in}{
\includegraphics[width=2.75in]{images/col_rnahelix.pdf} }
+\subsubsection{User Defined}
+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=2.1in]{images/col_user1.pdf}
+\includegraphics[width=2in]{images/col_user2.pdf}
+\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: Before you begin this exercise, ensure that the {\sl Apply Colour
\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.
+Clustal} 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}. Select a group
of around 4 similar sequences. Use the context menu (right click on the group)
{\bf See the video at: \url{http://www.jalview.org/training/Training-Videos}.}
}
-\subsubsection{User Defined}
-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=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).}
-\label{usercol}
-\end{center}
-\end{figure}
-
\exercise{User Defined Colour Schemes}{
-\exstep{Load a sequence alignment. Select the alignment menu option {\sl Colour $\Rightarrow$ User Defined}. A dialog window will open.}
+\label{colouex}
+\exstep{Load a sequence alignment. 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.
\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 {\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.
-\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 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}{
\begin{center}\centerline{
\includegraphics[width=2.5in]{images/mulv_tabs.pdf}}
\end{center}
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.
\begin{figure}[htbp]
% 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
+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}).
\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}.
+\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. }
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 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{Annotations can be reordered by clicking on the sequence 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 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 this icon up or down.}
-}
+\bf See the video at:
+\url{http://www.jalview.org/training/Training-Videos}.}
\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]{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]{3.5in}{\centerline{\includegraphics[width=2.8in]{images/image_eps.pdf}}
+\par \centerline{Zoom Detail of EPS image.}}
\subsubsection{PNG}
-\parbox[c]{3in}{
+\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=3in]{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.
$\Rightarrow$ 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}.
}
% left out for Glasgow 2016
analyse a given sequence or alignment and generate annotation for it.
-\section{Conservation, Quality and Consensus 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.
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
\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
+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.
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.
+\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, 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{Add an additional helix somewhere along the row by editing the file and
-re-importing it.
-
+(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{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.}
+function 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 for after you have completed exercise \ref{secstrpredex}:}
+\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
+Note: the
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.
+
\section{Importing Features from Databases}
\label{featuresfromdb}
Jalview supports feature retrieval from public databases.
-It includes built in parsers for Uniprot and ENA (or EMBL) records retrieved
+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
Sequence Fetcher} has access to. This function is accessed from the {\sl
Webservice $\Rightarrow$ 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, 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
{\sl ``Sequence not 100\% match'' dialog box}).
-\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.
-
\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.
The retrieved features are shown on the sequence and can be customised as described previously.
-\subsection{Colouring Features by Score or Description
-Text}
+\subsection{Customising Feature Display}
+
+\begin{figure}[htbp]
+\begin{center}
+\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}
+
+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 the feature's distribution or score attribute.
+These capabilities are described further in sections
+\ref{featureschemes} and \ref{featureordering}.
+
+
+\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}
% 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 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.
-
-\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 dialog 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.
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.
+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 dialog box appears, clicking the Sequence Feature
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 {\sl OK} or
-{\sl Cancel}.} }
+{\sl Cancel}.}
+\bf See the video at:
+\url{http://www.jalview.org/training/Training-Videos}.}
\chapter{Multiple Sequence Alignment}
\label{msaservices}
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 additonal sequences), but
+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]
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
\begin{figure}[htbc]
\center{
\includegraphics[width=3in]{images/jvaliwsparamsbox.pdf}
-\caption{{\bf Jalview's JABA alignment service parameter editing dialog box}.}
+\caption{{\bf An alignment service's parameter editing dialog box}.}
\label{jwsparamsdialog} }
\end{figure}
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}
the {\sl File $\Rightarrow$ Save As $\Rightarrow$ \ldots } submenu.
\exercise{Principal Component Analysis}
-{ \exstep{Load the alignment at
+{\label{pcaex}
+\exstep{Load the alignment at
\textsf{http://www.jalview.org/tutorial/alignment.fa}.}
-\exstep{Select the menu option {\sl Calculate $\Rightarrow$ Principal Component
-Analysis}.
-A new window will open. Move this window within the desktop so that the tree,
-alignment and PCA viewer windows are all visible.
+\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 the sequences on the
-alignment.
-} \exstep{ Select {\sl Calculate $\Rightarrow$ Calculate Tree $\Rightarrow$
-Neighbour Joining Using BLOSUM62}. A new tree window will appear.
-Place the mouse cursor on the tree window so that the
-tree partition location will divide the alignment into a number of groups, each of a different colour.
+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.}
{\bf See the video at:
\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.
{\sl View $\Rightarrow$ Mark Unlinked Leaves} menu option.
+
\begin{figure}
\begin{center}
\includegraphics[width=2.5in]{images/trees1.pdf}
\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}
+\begin{list}{$\circ$}{Ensure that you have at least 1G memory available in Jalview.}
+\item{Start with link:
+\url{http://www.jalview.org/services/launchApp?jvm-max-heap=1G}
+}
+\item{From
+{\href{http://www.jalview.org/development/development-builds}{the Development Builds}} in the Jalview web site, use
+the ``2G'' link in the``latest official build'' row and ``Webstart'' column.}
+\end{list}
+
+\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 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{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{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}
-{Ensure that you have at least 1G memory available in Jalview.
-
-{\sl (Start with 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 table, go to ``latest official build'' row and ``Webstart'' column, click
-on ``2G''.)}
-
-\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 tree window opens.}
-\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.}
-\exstep{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 by the 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. 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.
-{\bf See the video at:
-\url{http://www.jalview.org/training/Training-Videos}.}
-}
-\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}.
+%\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}.
-A warning dialog box {\bf ``Sequences not aligned'' } appears because the sequences input to the tree calculation are of different lengths. }
+%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}.
+%\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}.}
-}
+%{\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}, 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 into several sections.}
+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 using the calculated
-tree.
-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 within the View menu to aid navigation.}
+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).}
\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}
\exercise{Remove Redundant Sequences}{
-
+\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.}
}
\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 in the alignment window, select {\sl New View} to
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.
+\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}.}
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 in a structure viewer and the results interpreted on a 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}).
+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}
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 directed to the UCSF
-Chimera download page.
+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 can automatically determine which structures are associated with a
-sequence via its ID, and any associated database references. To do this, open
-the Sequence ID popup menu and select {\sl View 3D Structure}, to open the 3D
-Structure Chooser.
+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 attempt to discover identifiers for the sequence and from there discover any
-associated PDB structures. This can take a few seconds for each sequence and
-will be performed for all selected sequences. After this is done, you can see
-the added database references in a tool tip by mousing over the sequence
-ID\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.
+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}
\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 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]
\section{Viewing Structures}
-\label{viewAllStructures}
-The structure viewer is launched from the sequence ID context
-menu. To view a particular structure associated with a sequence in the
-alignment, select the sequence and right click the mouse to open context
-menu {\sl Structure $\Rightarrow$ 3D Structure data $\Rightarrow$} opens a Structure Chooser dialog box.
-The second way is most useful if you want to view all structural data available for
-a set of sequences in an alignment. Select all the sequence ids in the sequence
-ID panel and right click the mouse to open context
-menu {\sl Structure $\Rightarrow$ 3D Structure data $\Rightarrow$}. If any of
-the {\bf currently selected} sequences have structures associated they will
-appear in the Structure Chooser dialog box. The structures can be ranked by
-different parameters, 'Best Quality' is defaul option. Select the
-structures required and click {\sl View} to open a structure viewer containing
-the associated structures superposed according to the alignment.
-
-The structure to be displayed will be downloaded or loaded from
+\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
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}
automatic application of colour schemes when new structure data is added, or
when associated alignment views are modified.
-\exercise{Viewing Structures with the integrated Jmol
-Viewer}{\label{viewingstructex} \exstep{Load the alignment at
-\textsf{http://www.jalview.org/examples/exampleFile.jar}.}
+\exercise{Viewing Structures with the integrated Jmol Viewer}{
+\label{viewingstructex}
+\exstep{Load the alignment at
+\textsf{http://www.jalview.org/examples/exampleFile.jvp}.}
\exstep{Right-click on the
sequence ID label of {\sl FER1\_SPIOL} to open
the ID popup menu and select {\sl 3D Structure}. After a short pause, a
\url{http://www.jalview.org/training/Training-Videos}.}
}
-\exercise{Setting Chimera as the default 3D Structure Viewer}{\label{viewingchimera}
+\exercise{Aligning Structures using the Ferredoxin Sequence Alignment}{
+\label{superpositionex}
+
+\exstep{Continue with the Jalview project created in exercise
+\ref{viewingstructex}}
+
+\exstep{Open the 3D Structure chooser dialog from the popup menu for FER1\_MAIZE
+by right-clicking its ID (CMD-click on Macs), and selecting {\sl $\Rightarrow$
+3D Structure Data \ldots } }
+
+\exstep{Pick 1gaq from the Structure Chooser dialog, and make sure the {\sl Superpose} option is checked before clicking the {\bf Add}
+button to superimpose the structure associated with
+FER1\_MAIZE with the one associated with FER1\_SPIOL.
+
+{\sl 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, and hide all but columns 121
+through to 132 (you can do this via {\sl View $\Rightarrow$ Hide $\Rightarrow$
+All but selected region}).}
+\exstep{Select the newly created view in the {\sl Jmol $\Rightarrow$ Superpose
+With } submenu, and then recompute the superposition with {\sl Jmol
+$\Rightarrow$ Align 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.}
+
+\exstep{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).
+
+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.
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}.} }
+{\bf See the video at: \url{http://www.jalview.org/training/Training-Videos}.}}
+
\subsection{Superimposing Structures}
\label{superposestructs}
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}}
-
-\exstep{Open the 3D Structure chooser dialog from the popup menu for FER1\_SPIOL
-by right-clicking its ID (CMD-click on Macs), and selecting {\sl $\Rightarrow$
-3D Structure Data \ldots } }
-
-\exstep{Pick 1A70 from the Structure Chooser dialog, and click the {\bf View}
-button. 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 {\sl Yes}.
-
-{\sl 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, and hide all but columns 121
-through to 132 (you can do this via {\sl View $\Rightarrow$ Hide $\Rightarrow$
-All but selected region}).}
-\exstep{Select the newly created view in the {\sl Jmol $\Rightarrow$ Superpose
-With } submenu, and then recompute the superposition with {\sl Jmol
-$\Rightarrow$ Align 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.}
-
-\exstep{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).
-
-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
\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 1G of
-free memory available.
-
-{\sl See section \ref{memorylimits} for how to do this or click the following
-link:
-
-\url{http://www.jalview.org/services/launchApp?jvm-max-heap=2G} }}
\exstep{Retrieve the following PFAM alignments from the {\bf PFAM (full)} source
:
+
PF02008 PF01426 PF00145 (enter all three - they
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{Use the Find dialog to locate every DNMT1\_MOUSE sequence in the
-alignment and for each one, open the Structure Chooser via the ID popup
-menu ({\sl $\Rightarrow$ 3D Structure Data }. Select the DNMT1\_MOUSE.pdb
-structure from the `Cached Structures' view, and click {\bf View}.
+\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. For
+each one, 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 {\bf 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.}
sequences in the alignment.}
}
\exstep{Repeat the previous two steps for each of the other
-alignments. In each case, after selecting the DNMT1\_MOUSE.pdb structure and
-hitting the `View' button on the Structure Chooser dialog, Jalview will ask if you wish to create
-a new Jmol view. Respond {\bf `Yes'} each time. This will ensure ensure each sequence
-fragment is associated with the {\bf same} Jmol view. }
+ alignments. For those, after selecting the DNMT1\_MOUSE.pdb structure you
+ should select `Add' to ensure each domain alignment is associated
+ with the {\bf 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.
-{\sl The different shading schemes will allow regions of strong physicochemical conservation are
-highlighted on the domains in the structure.}
+{\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
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 Examine the regions strongly coloured at the interfaces betweeen each
+{\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 ? } }
+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.}
+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
prediction on the first sequence in the set (that is, the one that appears first in the alignment window).
\end{list}
-
-\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 JPred predictions. }
-\label{jpred}
-\end{center}
-\end{figure}
-
-
-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 JNet 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.
-
-
\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 wndow also provides options for
-reording and hiding autocalculated and sequence associated annotation. }
+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 $\Rightarrow$
{\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 JPred predictions. }
+\label{jpred}
+\end{center}
+\end{figure}
+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.
+
\section{Protein Disorder Prediction}
\label{protdisorderpred}
allows access to protein disorder prediction services provided by the configured
JABAWS servers.
-
\begin{figure}[htbp]
\begin{center}
\includegraphics[width=5in]{images/disorderpredannot.pdf}
\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.
highlight differences in disorder prediction across aligned sequences.
-\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}
-
\subsection{Navigating Large Sets of Disorder Predictions}
Figure \ref{alignmentdisorderannot} shows a single sequence annotated with
the associated sequence in the alignment display, and double clicking will
select that sequence.
-
\subsection{Disorder Predictors provided by JABAWS 2.0}
For full details of each predictor and the results that Jalview can display,
please consult
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
annotation panel and selecting \textbf{Show hidden annotation}.
\exercise{Protein Disorder Prediction}{
-%\label{protdispredex}
+\label{protdisorderex}
{\sl Before starting this exercise, make sure you enable the \protect{`Add
Temperature Factor'} option in your {\bf Structures} preferences. }
-\exstep{Open the alignment at:
+\exstep{Open the alignment from
\url{http://www.jalview.org/tutorial/interleukin7.fa}. }
-\exstep{Run the DisEMBL disorder predictor {\slvia} the {\slWeb Service
-$\Rightarrow$ Disorder Prediction } submenu.}
+\exstep{Run the DisEMBL disorder predictor {\sl via} the {\sl Web Service
+$\Rightarrow$ Disorder Prediction }.}
-\exstep{Select all the sequences, and open the Structure Chooser via the {\sl
-Sequence ID $\Rightarrow$ 3D Structure Data\ldots } popup menu. Hit the
-{\bf View} button to retrieve and show all PDB structures for the sequences.}
+\exstep{Select all the sequences. 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 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.
-
-{\sl Note how well the regions predicted to be disordered by the 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}.}
+\exstep{Apply the IUPred disorder prediction method. Tick the
+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 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.
+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.
\includegraphics[width=5in]{images/dnadasfeatures.pdf}
\caption{Uniprot and PDB sum features retrieved and mapped onto
-coding regions of EMBL record V00488 (an earlier version of Jalview is shown
+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 {\sl Wrapped} mode so the distinct exons can be seen.}
\exstep{Open the {\sl DAS Settings} tab in the {\sl Sequence Feature Settings\ldots}
}
\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.}
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