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+\usepackage[format=hang,margin=10pt]{caption}
+\usepackage{footnote}
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\usepackage{amssymb}
\usepackage{epstopdf}
\usepackage{hyperref}
+\usepackage{subfigure}
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-\title{Jalview 2.4: A manual and introductory tutorial }
-\author{David Martin, James Procter, Andrew Waterhouse, Geoff Barton}
+
+\title{Jalview 2.5: A manual and introductory tutorial }
+\author{David Martin, James Procter, Andrew Waterhouse and Geoff Barton}
+\date{Manual version 1.2.3 6th May 2011}
+
+\newcommand{\clearemptydoublepage}{\newpage{\pagestyle{empty}}\cleardoublepage}
+
% how the hell do we add another panel with text like : This tutorial introduces
% the user to the features of Jalview, a multiple sequence alignment editor and
% viewer available from http://www.jalview.org to the title page.
% \renewcommand{menustyle}{\tt} %do something more advanced here.
-\begin{document}
\newcounter{ecount}
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\renewcommand{\theecount} {\arabic{ecount}}
\renewcommand{\theexstep} {\arabic{ecount}.\alph{exstep}}
-\newcommand{\exercise}[2] { \refstepcounter{ecount}
+\newcommand{\exercise}[2] {
+\refstepcounter{ecount}
\begin{center} \fbox{\parbox[b][\height]{6in}{
-{\bf Exercise \theecount : #1 }
+{\bf
+% this doesn't work - page refs are off
+% \mbox{\addcontentsline{toc}{subsection}{ Exercise \theecount : #1 } }
+Exercise \theecount : #1 }
\par #2 }} \end{center}
-% this doesn't work - page refs are off \addcontentsline{toc}{subsection}{ Exercise \theecount : #1 }
+\pagebreak[0]
}
\newcommand{\exstep}[1]{ \stepcounter{exstep} {\sl \theexstep.} \begin{minipage}[t]{5.5in} #1 \end{minipage} \par \vspace *{1mm} }
%%% Remove the % on the next 2 lines to hide tutorials.
%\renewcommand{\exercise}[2]{}
%\renewcommand{\exstep}[1]{}
+\begin{document}
+
+
+\pagenumbering{}
+
+%\maketitle
+% we make our own title because JBP is not clever enough to work out how
+% to
+% do
+% this automatically
+
+\begin{center}
+
+{\Huge
+
+Jalview 2.7
+}
+\vspace{0.5in}
+{\huge
+
+A manual and introductory tutorial }
+
+\vspace{2.5in}
+
+{\large
+
+David Martin, James Procter, Andrew Waterhouse and Geoff Barton
+
+}
+
+\vspace{1.5in}
+
+College of Life Sciences, University of Dundee
+
+Dundee, Scotland DD1 5EH, UK
+
+
+\vspace{2in}
-\maketitle
-\tableofcontents
+Manual version 1.3.01
+
+6th December 2011
+
+
+\end{center}
+
+\newpage
+
+% \clearemptydoublepage
+
+% ($Revision$) 11th October 2010.}
+% TODO revise for 2.6
+
+\pagenumbering{roman}
+\setcounter{page}{1}
+\tableofcontents
+\clearemptydoublepage
+% \listoffigures
+% \newpage
+% \listoftables
+% \newpage
+\pagenumbering{arabic}
+\setcounter{page}{1}
\chapter{Basics}
+\label{jalviewbasics}
\section{Introduction}
\subsection{Jalview}
-Jalview is a sequence multiple alignment viewer, editor and analysis tool.
+Jalview is a multiple sequence alignment viewer, editor and analysis tool.
Jalview is designed to be platform independent (running on Mac, MS Windows, Linux
and any other platform that supports Java), capable of editing and analysing
large alignments (thousands of sequences) with minimal degradation in
FASTA, Clustal, MSF(GCG) and PIR.
-There are two types of Jalview program. The Jalview Desktop is a stand alone
-application that provides powerful editing, visualization, annotation and
-analysis capabilities. The JalviewLite applet has the same core visualization,
-editing and analysis capabilities as the desktop, but is designed to be embedded
-in a web page, allowing customised integration with alignment databases such as
-pFam\footnote{A demonstration version of Jalview (Jalview Micro Edition) also
-runs on a mobile phone but the functionality is limited to sequence colouring.}.
+There are two types of Jalview program. The {\bf Jalview Desktop} is a stand
+alone application that provides powerful editing, visualization, annotation and
+analysis capabilities. The {\bf JalviewLite} applet has the same core
+visualization, editing and analysis capabilities as the desktop, without the
+desktop's webservice and figure generation capabilities. It is designed to be
+embedded in a web page\footnote{A demonstration version of Jalview (Jalview Micro
+Edition) also runs on a mobile phone but the functionality is limited to sequence
+colouring.}, to allow customisable display of alignments for web sites such as
+{\bf pfam}\footnote{\url{http://pfam.sanger.ac.uk}}.
-Jalview 2.4 was released in September 2008. The Jalview Desktop in this version
+Jalview 2.7 was released in September 2011. The Jalview Desktop in this version
provides access to sequence, alignment and protein structure databases, and
-alignment and analysis web services, and includes the Jmol\footnote{ Provided
-under the LGPL licence at http://www.jmol.org} protein structure viewer. It is
-also a Distributed Annotation System (DAS) client\footnote{with thanks to Andreas
-Prlic} which facilitates the retrieval and display of third party sequence
-annotation in association with sequences and any associated structure.
+includes the Jmol\footnote{ Provided under the LGPL licence at
+\url{http://www.jmol.org}} protein structure viewer. It includes a
+Distributed Annotation System (DAS) client\footnote{with thanks to Andreas Prlic} which facilitates the retrieval and display of third party sequence
+annotation in association with sequences and any associated structure. It also
+provides a graphical user interface for {\bf Ja}va {\bf B}ioinformatics
+{\bf A}nalysis {\bf W}eb {\bf S}ervices (JABAWS) which enable access to a
+range of multiple sequence alignment programs.
\subsection{Jalview's Capabilities}
% TODO add references to appropriate sections for each capability described here.
-Figure \ref{jvcapabilities} gives an overview of the main features of the Jalview desktop application. Its primary function is the editing and visualization of sequence alignments, and their interactive analysis. Tree building, principle components analysis, physico-chemical property conservation and sequence consensus analyses are built in to the program. Web services enable Jalview to access remote alignment and secondary structure prediction services, as well as to retrieve protein and nucleic acid sequences, alignments, protein structures and sequence annotation. Sequences, alignments, trees, structures, features and alignment annotation may also be exchanged with the local filesystem. Multiple visualizations of an alignment may be worked on simultaneously, and the user interface provides a comprehensive set of controls for colouring and layout. Alignment views are dynamically linked with Jmol structure displays, a tree viewer and spatial cluster display, facilitating interactive exploration of alignment structure. Finally, the application provides a Jalview project format enabling the current state of an alignment and analysis windows to be stored, and WYSIWIG style figure generation capabilities for use in publications.
+Figure \ref{jvcapabilities} gives an overview of the main features of the Jalview desktop application. Its primary function is the editing and visualization of sequence alignments, and their interactive analysis. Tree building, principal components analysis, physico-chemical property conservation and sequence consensus analyses are built in to the program. Web services enable Jalview to access remote alignment and secondary structure prediction programs, as well as to retrieve protein and nucleic acid sequences, alignments, protein structures and sequence annotation. Sequences, alignments, trees, structures, features and alignment annotation may also be exchanged with the local filesystem. Multiple visualizations of an alignment may be worked on simultaneously, and the user interface provides a comprehensive set of controls for colouring and layout. Alignment views are dynamically linked with Jmol structure displays, a tree viewer and spatial cluster display, facilitating interactive exploration of the alignment's structure. The application provides its own Jalview project file format in order to store the current state of an alignment and analysis windows. Jalview also provides WYSIWIG\footnote{WYSIWIG: What You See Is What You Get.} style figure generation capabilities for the preparation of alignments for publication.
\begin{figure}[htbp]
\begin{center}
-\includegraphics[width=5.5in]{images/jvcapabilities.pdf}
+\includegraphics[width=5.8in]{images/jvcapabilities.pdf}
+\caption{{\bf Capabilities of the Jalview Desktop.} The Jalview Desktop Application provides a stable environment for the creation, editing and analysis of alignments and the generation of figures.}
\label{jvcapabilities}
\end{center}
-\caption{{\bf Capabilities of the Jalview Desktop.} The Jalview Desktop Application provides a stable environment for the creation, editing and analysis of alignments and the generation of figures.}
\end{figure}
\subsubsection{Jalview History}
Searle and Geoff Barton at the University of Oxford and then the European
Bioinformatics Institute. Development of Jalview 2 was made possible with
eScience funding from the BBSRC\footnote{Biotechnology and Biological Sciences
-Research Council grant {\sl "VAMSAS: Visualisation and Analysis of Molecules,
+Research Council grant {\sl ``VAMSAS: Visualization and Analysis of Molecules,
Sequence Alignments and Structures"}, a joint project to enable interoperability
between Jalview, TOPALi and AstexViewer.} in 2004, enabling Andrew Waterhouse and
-Jim Procter to re-engineer the original program to introduce current developments
-in bioinformatics and take advantage of the latest web and Java technology. The
-2.4 version released in September 2008 introduces experimental support for a data
-exchange framework developed as part of this project, to enable Jalview to be
-used in conjunction with other programs for the analysis of sequences, alignments
-and structures. Jalview continues to be one of the worlds most
-popular\footnote{and in the authors opinion, the best.} sequence alignment and
-analysis tools.
+Jim Procter to re-engineer the original program to introduce contemporary developments
+in bioinformatics and take advantage of the latest web and Java technology.
+Jalview's development is now supported for a further 5 years from October 2009
+by an award from the BBSRC's Tools and Resources fund. In 2010 and 2011, Jalview
+benefitted from the \href{http://code.google.com/soc/}{Google Summer of Code},
+when Lauren Lui and Jan Engelhardt introduced new features for handling RNA
+alignments and secondary structure annotation.
+
+
+%TODO describe future plans in history ? not a good idea.
+% Jalview continues to be one of the worlds most popular\footnote{and in the authors opinion, the best.} sequence alignment and analysis tools.
\subsubsection{Citing Jalview}
-If you use Jalview in your work you should cite {\sl "The Jalview Java alignment
-editor"} Michele Clamp, James Cuff, Stephen M. Searle and Geoffrey J. Barton
-(2004) {\sl Bioinformatics} {\bf 20} 426-427.
+If you use Jalview in your work you should cite:\newline
+{\sl ``Jalview Version 2 - a multiple sequence alignment editor and analysis workbench"}\newline
+Waterhouse, A.M., Procter, J.B., Martin, D.M.A, Clamp, M. and Barton, G. J. (2009) \newline {\sl Bioinformatics} doi: 10.1093/bioinformatics/btp033
+
+This paper supersedes the original Jalview publication:\newline {\sl ``The Jalview Java alignment
+editor"} \newline Michele Clamp, James Cuff, Stephen M. Searle and Geoffrey J. Barton (2004) \newline {\sl Bioinformatics} {\bf 20} 426-427.
+
\r\subsection{About this tutorial }
This tutorial is written in a manual format with short exercises where
-appropriate, typically at the end of each section. The first chapter concerns the
+appropriate, typically at the end of each section. This chapter concerns the
basic operation of Jalview and should be sufficient for those who just want to
-open an alignment, perform basic editing and colouring, and produce publication
-and presentation quality graphical output.
-
-The second chapter covers analysis in Jalview, sequence alignment and
-visualisation of external features and structural data.
-
-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
-Jalview experience.
+load Jalview (Section \ref{startingjv}), open an alignment (Section \ref{loadingseqs}), perform basic editing and colouring (Section \ref{selectingandediting} and Section \ref{colours}), and produce publication
+and presentation quality graphical output (Section \ref{layoutandoutput}).
+
+Chapter \ref{analysisannotation} covers the additional visualization and
+analysis techniques that Jalview provides. This includes working with the
+embedded Jmol molecular structure viewer, building and viewing trees and PCA
+plots, and using trees for sequence conservation analysis. An overview of
+the Jalview Desktop's webservices is given in Section \ref{jvwebservices}, and
+the alignment and secondary structure prediction services are described
+in detail in Sections \ref{msaservices} and \ref{protsspredservices}. Following
+this, Section \ref{featannot} details the creation and visualization of sequence
+and alignment annotation, and the retrieval of sequences and annotation from
+databases and DAS Servers. Finally, Section \ref{workingwithnuc} discusses
+specific features of use when working with nucleic acid sequences and protein
+coding regions.
+
+%^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
+%Jalview experience.
\subsubsection{Typographic Conventions}
-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). Menu options are given as a path from the menu 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.
+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). Menu options are given as a path from the menu 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.
\r\section{Obtaining and starting The Jalview Desktop Application}
+\label{startingjv}
\begin{figure}[htbp]
\begin{center}
\includegraphics[width=4.5in]{images/download.pdf}
This tutorial is based on the application version of Jalview, the Jalview
Desktop. Much of the information will also be useful for users of the JalviewLite
-applet. This version of Jalview 2 is designed to be launched from web pages, but
-has the same core editing, analysis and visualization capabilities (see the
+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
includes additional support for interaction with external web services, and
(http://www.jalview.org/).
Jalview can be started directly with webstart by navigating to the Download page
-(via the menu on the left hand side), and clicking the 'Start with Java Webstart'
-button. (figure \ref{download}). This will always launch the latest stable
+(via the menu on the left hand side), and clicking the `Start with Java Webstart'
+button. (Figure \ref{download}). This will always launch the latest stable
release of Jalview.\par
The application will start automatically though you may be prompted to accept a
-security certificate signed by the Barton Group. You can trust us so click trust
-or accept as appropriate. The splash screen (figure \ref{splash}) gives
-information about the latest version and the paper to cite in your publications.
-This information is also available on the Jalview web site and from the {\sl Help
-$\Rightarrow$ About} menu option.
+security certificate signed by the Barton Group. You can always trust us, so click trust
+or accept as appropriate. The splash screen (Figure \ref{splash}) gives
+information about the version and build date that you are running,
+information about later versions (if available), and the paper to cite in your
+publications. This information is also available on the Jalview web site and from the {\sl Help $\Rightarrow$ About} menu option.
%[fig 2]
\begin{figure}[htbp]
\end{center}
\end{figure}
\rWhen Jalview starts it will automatically load an example alignment from the
-Jalview site. Later in the tutorial we will discover how to alter this behaviour.
-This alignment looks like figure \ref{startpage}.
+Jalview site. This behaviour can be changed in the Jalview Desktop preferences
+dialog opened from the Desktop's {\sl Tools $\Rightarrow$ Preferences..} menu.
+This alignment will look like the one in Figure \ref{startpage} (this is taken
+from Jalview version 2.7).
%[figure 3 ]
\begin{figure}[htbp]
\end{figure}
+\subsubsection{Jalview News RSS Feed}
+
+From time to time, important announcements are made available to users of the
+Jalview Destop {\sl via} the Jalview News reader. This window will open
+automatically when new news is available, and can also be accessed via the
+Desktop's `{\sl Tools $\Rightarrow$ Show Jalview News}' menu entry.
+
+\begin{figure}[htbp]
+\begin{center}
+\includegraphics[height=3in]{images/jvrssnews.pdf}
+\caption{{\bf The Jalview News Reader.} The news reader opens automatically
+when new articles are available from the Jalview Desktop's news channel.}
+\label{jalviewrssnews}
+\end{center}
+\end{figure}
+
+
\exercise{Starting Jalview}{
\label{start}
-\exstep{Point your web browser at the \href{http://www.jalview.org}{Jalview web site} and start Jalview by clicking on the 'Start with Java WebStart' button.} }
+\exstep{Point your web browser at the \href{http://www.jalview.org}{Jalview web site} and start Jalview by clicking on the `Start with Java WebStart' button.}
+\exstep{Open the Jalview Desktop's user preferences dialog (from the Tools
+menu), and untick the checkbox adjacent to the `Open file' entry in the
+`Visual' preferences tab.}
+\exstep{Click OK to save the preferences, then \em{launch another Jalview
+instance from the web site}. The example alignment should not be
+loaded when the new Jalview instance starts up.}
+{\sl Note: Should you want to reload the example alignment, then select the
+{\em File$\Rightarrow$ From URL} entry from the Desktop menu, and click on the
+URL history button on the right hand side of the dialog box that opens to
+recover the example file's URL, followed by OK, to open the file. } }
\subsection{Getting Help}
-
+\label{gettinghelp}
\subsubsection{Built in documentation}
-Jalview has comprehensive on-line help documentation. Select {\sl Help $\Rightarrow$ Documentation} from the main window menu and a new window will open (Figure \ref{help}). The appropriate topic can then be selected from the 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.
+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 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.
\begin{figure}[htbp]
\subsubsection{Email lists}
-The Jalview Discussion list {\tt jalview-discuss@jalview.org} provides a forum for Jalview users and developers to raise problems and exchange ideas. The Jalview developers may also be contacted directly in the case of specific bugs, feature requests and so on via the email address {\tt help@jalview.org}. Finally, the {\tt jalview-announce@jalview.org} list can be subscribed to if you wish to be kept informed of new releases and developments. Archives and mailing list subscription details can be found on the Jalview web site.
+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 kept informed of new releases and developments. Archives and mailing list subscription details can be found on the Jalview web site.
\r\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: normal mode, where editing and navigation is performed using the mouse, and cursor mode where editing and navigation are performed using the keyboard. The F2 key is used to switch between these two modes.
\end{figure}
\subsection{Navigation in Normal mode}
-\rJalview always starts up in Normal mode, where the mouse is used to interact with the displayed alignment view. You can move about the alignment by clicking and dragging the ruler scroll bar to move horizontally, or by clicking and dragging the alignment scroll bar to the right of the alignment to move vertically. If all the rows or columns in the alignment are displayed, the scroll bars will not be visible.\r\r 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 visualise 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 window menu (Figure \ref{overview}).\r%(figure 4)
+\rJalview always starts up in Normal mode, where the mouse is used to interact with the displayed alignment view. You can move about the alignment by clicking and dragging the ruler scroll bar to move horizontally, or by clicking and dragging the alignment scroll bar to the right of the alignment to move vertically. If all the rows or columns in the alignment are displayed, the scroll bars will not be visible.\r\r 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 window menu (Figure \ref{overview}).\r%(Figure4)
\begin{figure}[htbp]
\begin{center}
\includegraphics[width=4.5in]{images/overview.pdf}
\label{overview}
\end{center}
\end{figure}
-\rThe red box in the overview window shows the current view in the alignment window. A percent identity histogram is plotted below the alignment overview. Shaded parts indicate rows and columns of the alignment that are hidden (in this case, a single row at the bottom of the alignment). You can navigate around the alignment by dragging the red box. %Try this now and see how the view in the alignment window changes.\r\r\parbox[c]{3in}{Alignment and analysis windows are closed by clicking on the usual 'close' icon (indicated by arrows on Mac OS X). If you want to close all the alignments and analysis windows at once, then use the {\sl Window $\Rightarrow$ Close All} option from the Jalview desktop (\em{warning: make sure you have saved your work because this cannot be undone !}). }
+\rThe red box in the overview window shows the current view in the alignment window. A percent identity histogram is plotted below the alignment overview. Shaded parts indicate rows and columns of the alignment that are hidden (in this case, a single row at the bottom of the alignment - see Section \ref{hidingregions}). You can navigate around the alignment by dragging the red box. %Try this now and see how the view in the alignment window changes.\r\r\parbox[c]{3in}{Alignment and analysis windows are closed by clicking on the usual `close' icon (indicated by arrows on Mac OS X). If you want to close all the alignments and analysis windows at once, then use the {\sl Window $\Rightarrow$ Close All} option from the Jalview desktop (\em{warning: make sure you have saved your work because this cannot be undone !}). }
\parbox[c]{3in}{\centerline{\includegraphics[width=2.5in]{images/start_closeall.pdf}
}}
\subsection{Navigation in Cursor mode}
-
+\label{cursormode}
\parbox[c]{5in}{Cursor mode navigation enables the experienced user to quickly and precisely navigate, select and edit parts of an alignment. On pressing F2 to enter cursor mode the position of the cursor is indicated by a black background and white text. The cursor can be placed using the mouse or moved by pressing the arrow keys ($\uparrow$, $\downarrow$, $\leftarrow$, $\rightarrow$).
}\parbox[c]{1.25in}{\centerline{\includegraphics[width=0.8in]{images/cursor1.pdf}}}
Move to sequence 7 by pressing {\sl 7 S}. Move to column 18 by pressing {\sl 1 8 C}. Move to residue 18 by pressing {\sl 1 8 P}. Note that these can be two different positions if gaps are inserted into the sequence. Move to sequence 5, column 13 by typing {\sl 1 3 , 5 [RETURN]}.
}
}
-\r\section{Loading your own sequences}\r\rJalview 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.\r\r\subsection{Drag and Drop}\r In some operating systems (Mac OS X, Windows XP) you can just drag a file icon from a file browser window and drop it on an open Jalview application window. The file will then be opened as a new alignment window. %You can try this with the tutorial file you have downloaded. When you have opened the file, close it again by selecting the close control on the window.\rIf you drop an alignment file onto an open alignment window it will be appended to that alignment.\r% (Figure \ref{drag})
+\subsection{The Find Dialog Box}
+\label{searchfunction}
+A further option for navigation is to use the {\sl Select $\Rightarrow$
+Find\ldots} function. This opens a dialog box into which can be entered regular
+expressions for searching sequences and sequence IDs, or sequence numbers. Hitting the [Find next] button will highlight the first (or next) occurrence of that pattern in the sequence ID panel or the alignment, and will adjust the view in order to display the highlighted region. The Jalview help provides comprehensive documentation for this function, and a quick guide to the regular expressions that can be used with it.
+
+%TODO insert a figure for the Find dialog box
+
+\r\section{Loading your own sequences}\r\label{loadingseqs}\rJalview 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.\r\r\subsection{Drag and Drop}\r In most operating systems you can just drag a file icon from a file browser\r 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\rto that alignment. Drag and drop also works when loading data from a URL -\rsimply drag the link or url from the address panel of your browser on to an\ralignment or the Jalview desktop background and Jalview will load data from the\rURL directly.\r% (Figure \ref{drag})
% %[fig 5]\r% \begin{figure}[htbp]\r% \begin{center}\r% \includegraphics[width=2in]{images/drag2.pdf}\r% \includegraphics[width=2in]{images/drag3.pdf}\r% \caption{{\bf An alignment can be opened by dragging the file onto the Jalview window. }}\r% \label{drag}\r% \end{center}\r% \end{figure}\r% \r% %\includegraphics[width=2in]{images/drag1.pdf}
-\r\subsection{From a File}\r Jalview can read sequence alignments from a sequence alignment file. This is a text file, not a word processor document. For entering sequences from a wordprocessor document see Cut and Paste (section \ref{cutpaste}) below. Select {\sl File $\Rightarrow$ Input Alignment $\Rightarrow$ From File} from the main menu (Figure \ref{loadfile}). You will then get a file selection window where you can choose the file to open. Remember to select the appropriate file type. Jalview can automatically identify some sequence file formats.\r\r%[fig 6]\r\begin{figure}[htbp]
+\r\subsection{From a File}\r Jalview can read sequence alignments from a sequence alignment file. This is a text file, not a word processor document. For entering sequences from a wordprocessor document see Cut and Paste (Section \ref{cutpaste}) below. Select {\sl File $\Rightarrow$ Input Alignment $\Rightarrow$ From File} from the main menu (Figure \ref{loadfile}). You will then get a file selection window where you can choose the file to open. Remember to select the appropriate file type. Jalview can automatically identify some sequence file formats.\r\r%[fig 6]\r\begin{figure}[htbp]
\begin{center}
\includegraphics[width=2in]{images/loadfilebox.pdf}
%\includegraphics[width=2in]{images/loadfilebox2.pdf}
\label{loadfile}
\end{center}
\end{figure}
-\r\subsection{From a URL}\r Jalview can read sequence alignments directly from a URL. Please note that the files must be in a sequence alignment format - a pretty HTML alignment or graphics file cannot be read by Jalview. \rSelect {\sl File $\Rightarrow$ Input Alignment $\Rightarrow$ From URL} from the main menu and a window will appear asking you to enter the URL (figure \ref{loadurl}). Jalview will attempt to automatically discover the file format.\r\r%[fig 7]
+\r\subsection{From a URL}\r Jalview can read sequence alignments directly from a URL. Please note that the files must be in a sequence alignment format - a pretty HTML alignment or graphics file cannot be read by Jalview. \rSelect {\sl File $\Rightarrow$ Input Alignment $\Rightarrow$ From URL} from the main menu and a window will appear asking you to enter the URL (Figure\ref{loadurl}). Jalview will attempt to automatically discover the file format.\r\r%[fig 7]
\begin{figure}[htbp]
\begin{center}
\includegraphics[width=2in]{images/menuloadurl.pdf}
\end{center}
\end{figure}
\r\subsection{Cut and Paste}
-\label{cutpaste}\rDocuments such as those produced by Microsoft Word cannot be readily understood by Jalview. The way to read sequences from these documents is to select the data from the document and copy it to the clipboard. Select {\sl File $\Rightarrow$ Input Alignment $\Rightarrow$ From Textbox} from the main menu and a textbox window will appear (Figure \ref{loadtext}). Paste the sequences you have copied to the clipboard and, presuming that they are in the right format, Jalview will happily read them.\r%[fig 8]
+\label{cutpaste}\rDocuments such as those produced by Microsoft Word cannot be readily understood\rby Jalview. The way to read sequences from these documents is to select the\rdata from the document and copy it to the clipboard. There are two ways to\rdo this. One is to right-click on the desktop background, and select the\r`Paste to new alignment' option in the menu that appears. The other is to select\r{\sl File $\Rightarrow$ Input Alignment $\Rightarrow$ From Textbox} from the\rmain menu, and paste the sequences into the textbox window that will appear\r(Figure \ref{loadtext}). In both cases, presuming that they are in the right\rformat, Jalview will happily read them into a new alignment window.\r%[fig 8]
\begin{figure}[htbp]
\begin{center}
\end{figure}
\r\subsection{From a public database}
-\label{fetchseq}\rJalview can retrieve sequences and sequence alignments from the public databases housed at the European Bioinformatics Institute, such as Uniprot, Pfam and the PDB, as well as any DAS sequence server registered at the configured DAS registry. This facility avoids having to manually locate, save and load the sequences, and 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}). Use the menu box to select the appropriate database, enter a sequence ID/accession number, or several separated by a semicolon and Jalview will attempt to retrieve it/them from the chosen database source. Example queries are provided to test that a source is operational, but can also be used as a guide on the type of accession numbers understood by the source.\r%[fig 9]\r\begin{figure}[htbp]
+\label{fetchseq}\rJalview can retrieve sequences and sequence alignments from the public
+databases housed at the European Bioinformatics Institute, such as Uniprot,\rPfam and the PDB, as well as any DAS sequence server registered at the\rconfigured DAS registry. This facility avoids having to manually locate, save\rand load the sequences, and allows Jalview to gather additional metadata\rprovided by the source, such as annotation and database cross references.\rSelect {\sl File $\Rightarrow$ Fetch Sequence(s) \ldots} from the main menu and\ra window will appear (Figure \ref{loadseq}). Use the menu box to select the\rappropriate database, enter a sequence ID/accession number, or several\rseparated by a semicolon and Jalview will attempt to retrieve it/them from the
+chosen database source. Example queries are provided to test that a source is\roperational, and can also be used as a guide for the type of accession numbers\runderstood by the source.\r%[fig 9]\r\begin{figure}[htbp]
\begin{center}
\includegraphics[width=2.5in]{images/fetchseq.pdf}
\caption{{\bf Retrieving sequences from a public database}}
\label{loadseq}
\end{center}
\end{figure}
+
\exercise{Loading sequences}{
\label{load}
\exstep{Start Jalview then close all windows by selecting {\sl Window $\Rightarrow$ Close All} from the main menu}
\exstep{Select {\sl File $\Rightarrow$ Input Alignment $\Rightarrow$ From URL} from the main menu and enter \textsf{http://www.jalview.org/tutorial/alignment.fa} in the box. Click {\sl OK} and the alignment should load.
}
-\exstep{Close all windows using the {\sl Window $\Rightarrow$ Close All} main menu option. Point your web browser to \textsf{http://www.jalview.org/tutorial/alignment.fa} and save the file to your desktop. Open this file in Jalview by selecting {\sl File $\Rightarrow$ Input Alignment $\Rightarrow$ From File} from the main menu and browsing to the appropriate location. Click OK and load the alignment
-}
-\exstep{
-Drag the alignment.fa file from the desktop onto the Jalview window. The alignment should open. Try dragging onto an empty Jalview and onto an existing alignment and observe the results.
-}
-\exstep{
-Select {\sl File $\Rightarrow$ Fetch Sequence(s).. } from the main menu. Select the {\sl PFAM (seed)} database and enter the accession number PF03460. Click OK. An alignment of about 107 sequences should load.
-}
-\exstep{Open the URL \textsf{http://www.jalview.org/tutorial/alignment.fa} in a web browser. Select and copy the entire text to the clipboard (usually via the browser's {\sl Edit $\Rightarrow$ Copy} menu option). Ensure Jalview is running and 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 Close} and the alignment will be loaded.
-}
-}\r\section{Writing sequence alignments}\r\subsection{Saving the alignment}\rJalview allows the current sequence alignments to be saved to file so they can be restored at a later date, passed to colleagues or analysed in other programs. From the alignment window menu select {\sl File $\Rightarrow$ Save As} and a dialog box will appear (Figure \ref{savealign}). You can navigate to an appropriate directory in which to save the alignment. Jalview will remember the last filename and format used to save (or load) the alignment, enabling you to quickly update the file after editing by using the {\sl File $\Rightarrow$ Save} entry.
-
-Jalview offers several different formats in which an alignment can be saved. The jalview format is the only one which will preserve the colours, groupings and similar information in the alignment. The other formats produce text files containing just the sequences with no visualisation information, although some allow limited annotation and sequence features to be stored (e.g. AMSA). Unfortunately only Jalview can read Jalview files. The {\sl File $\Rightarrow$ Output To Textbox} menu option allows the alignment to be copied and pasted into other documents or web servers.
+\exstep{Close all windows using the {\sl Window $\Rightarrow$ Close All} main
+menu option. Point your web browser at the same \href{http://www.jalview.org/tutorial/alignment.fa}{URL} and save the file to
+your desktop. Open this file in Jalview by selecting {\sl File $\Rightarrow$ Input Alignment $\Rightarrow$ From File} from the main menu and browsing to the appropriate location. Click OK and load the alignment }
+\exstep{Drag the alignment.fa file from the desktop onto the Jalview window. The
+alignment should open. Try dragging onto an empty Jalview and onto an existing
+alignment and observe the results. You can also try dragging the
+\href{http://www.jalview.org/tutorial/alignment.fa}{URL} directly onto Jalview.
+}
+\exstep{ Select {\sl File $\Rightarrow$ Fetch Sequence(s).. } from the main menu. Select the {\sl PFAM (seed)} database and enter the accession number PF03460. Click OK. An alignment of about 107 sequences should load. }
+\exstep{Open the \href{http://www.jalview.org/tutorial/alignment.fa}{URL} in a
+web browser. Select and copy the entire text to the clipboard (usually via the
+browser's {\sl Edit $\Rightarrow$ Copy} menu option). Ensure Jalview is running
+and 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. } }
+
+\subsection{Memory Limits}
+\label{memorylimits}
+Jalview is a Java program. One unfortunate implication of this is that it does
+not allow Jalview to dynamically request additional memory from the operating
+system. It is important, therefore, that you ensure that you have allocated
+enough memory to work with your data. On most occasions, Jalview will warn you
+when you have tried to load an alignment that is too big to fit in to memory (for
+instance, some of the PFAM alignments are {\bf very} large). You can find out how
+much memory is available to Jalview with the desktop window's {\sl $\Rightarrow$
+Tools $\Rightarrow$ Show Memory Usage} function, which enables the display of the
+currently available memory at the bottom left hand side of the Desktop window's
+background. Should you need to increase the amount of memory available to
+Jalview, full instructions are given in both the built in documentation and on
+the JVM memory parameters page (http://www.jalview.org/jvmmemoryparams.html) on
+the website.
+
+\r\section{Writing sequence alignments}
+\label{savingalignments} \subsection{Saving the alignment} Jalview allows the
+current sequence alignments to be saved to file so they can be restored at a
+later date, passed to colleagues or analysed in other programs. From the
+alignment window menu select {\sl File $\Rightarrow$ Save As} and a dialog box
+will appear (Figure \ref{savealign}). You can navigate to an appropriate
+directory in which to save the alignment. Jalview will remember the last filename
+and format used to save (or load) the alignment, enabling you to quickly update
+the file after editing by using the {\sl File $\Rightarrow$ Save} entry.
+
+Jalview offers several different formats in which an alignment can be saved. The jalview format is the only one which will preserve the colours, groupings and similar information in the alignment. The other formats produce text files containing just the sequences with no visualization information, although some allow limited annotation and sequence features to be stored (e.g. AMSA). Unfortunately only Jalview can read Jalview files. The {\sl File $\Rightarrow$ Output To Textbox} menu option allows the alignment to be copied and pasted into other documents or web servers.
\r%[fig 10]
\begin{figure}[htbp]
\begin{center}
\end{center}
\end{figure}
-\subsection{Jalview Projects}\r\parbox[c]{4in}{If you wish to save the complete Jalview session rather than just one alignment (e.g. because you have calculated trees or multiple different alignments) then your work should be saved as a Jalview Project file. From the main menu select {\sl File $\Rightarrow$ Save Project} and a file save dialog box will appear. Loading a project will restore Jalview to exactly the view at which the file was saved, complete with all alignments, trees, annotation and displayed structures rendered appropriately.}
+\subsection{Jalview Projects}\r\parbox[c]{4in}{If you wish to save the complete Jalview session rather than just one alignment (e.g. because you have calculated trees or multiple different alignments) then your work should be saved as a Jalview Project file\footnote{Tip: Ensure that you have allocated plenty of memory to Jalview when working with large alignments in Jalview projects. See Section \protect \ref{memorylimits} above for how to do this.}.
+From the main menu select {\sl File $\Rightarrow$ Save Project} and a file save dialog box will appear. Loading a project will restore Jalview to exactly the view at which the file was saved, complete with all alignments, trees, annotation and displayed structures rendered appropriately.
+}
\parbox[c]{2in}{
\centerline {
\includegraphics[width=1.5in]{images/saveproj.pdf}
\exercise{Saving Alignments}{
\label{save}
-\exstep{Start Jalview, close all windows and load the ferrodoxin alignment from pFam (accession number PF03460 (see Exercise \ref{load}).
-}
+\exstep{Start Jalview, close all windows and load the ferredoxin alignment from
+PFAM (PFAM Seed accession number PF03460 (see Exercise \ref{load}). }
\exstep{
Select {\sl File $\Rightarrow$ Save As} from the alignment window menu. Choose a location into which to save the alignment and select a format. All formats except {\sl Jalview } can be viewed in a normal text editor (e.g. Notepad) or in a web browser. Enter a file name and click {\sl Save}. Check this file by browsing to it with your web browser or by closing all windows and opening it with Jalview.
}
\exstep{ Repeat the previous step trying different file formats.}
\exstep{Select {\sl File $\Rightarrow$ Output to Textbox $\Rightarrow$ FASTA}. You can select and copy this alignment to the clipboard using the textbox menu options {\sl Edit $\Rightarrow$ Select All} followed by {\sl Edit $\Rightarrow$ Copy}. The alignment can then be pasted into any application of choice, e.g. a word processor or web form.
}
-\exstep{Ensure at least one alignment window is shown in Jalview. Open the overview window and scroll to any part of the alignment. Select {\sl File $\Rightarrow$ Save Project} from the main menu and save in a suitable place. Close all windows and then load the project via the {\sl File $\Rightarrow$ Save Project} menu option. Note how all the windows and positions are exactly as they were when they were saved.
-}
-}
-\r\r\section{Selecting and editing sequences} \rJalview makes extensive use of selections which are arbitrary regions in an alignment. This section illustrates how to make and use selections and groups.
+\exstep{Ensure at least one alignment window is shown in Jalview. Open the
+overview window and scroll to any part of the alignment. Select {\sl File
+$\Rightarrow$ Save Project} from the main menu and save in a suitable place.
+Close all windows and then load the project via the {\sl File $\Rightarrow$ Load Project} menu option. Note how all the windows and positions are exactly as they were when they were saved. } }
+\r\r\section{Selecting and editing sequences}
+\label{selectingandediting} \rJalview makes extensive use of selections - most of the commands available from
+its menus operate on the currently selected region of the alignment, either to
+change their appearance or perform some kind of analysis. This section
+illustrates how to make and use selections and groups.
\r\subsection{Selecting parts of an alignment}
Selections can be of arbitrary regions in an alignment, one or more complete columns, or one or more complete sequences.
-A selected region can be copied and pasted as a new alignment using the {\sl Edit $\Rightarrow$ Copy} and {\sl Edit $\Rightarrow$ Paste $\Rightarrow$ As New Alignment} alignment window menu options.
+A selected region can be copied and pasted as a new alignment using the {\sl Edit $\Rightarrow$ Copy} and {\sl Edit $\Rightarrow$ Paste $\Rightarrow$ New Alignment} alignment window menu options.
To clear (unselect) the selection press the [ESC] (escape) key.
\end{center}
\end{figure}
-\subsubsection{Selecting columns}\rTo select the same residues in all sequences, click and drag along the alignment ruler. This selects the entire height of the alignment. Ranges of positions can also be selected by clicking on the first position then holding down the [SHIFT] key whilst clicking the other end of the selection. Discontinuous regions can be selected by holding down [CTRL] and clicking on positions to add to the selection. Selected columns are indicated by red highlighting in the ruler bar (Figure \ref{selectcols}).\r%[fig 13]
+\subsubsection{Selecting columns}\rTo select the same residues in all sequences, click and drag along the alignment ruler. This selects the entire height of the alignment. Ranges of positions can also be selected by clicking on the first position then holding down the [SHIFT] key whilst clicking the other end of the selection. Discontinuous regions can be selected by holding down [CTRL] and clicking on positions to add to the column selection. Note that each [CTRL]-Click changes the current selected sequence region to that column, but adds to the column selection. Selected columns are indicated by red highlighting in the ruler bar (Figure \ref{selectcols}).\r%[fig 13]
\begin{figure}[htbp]
\begin{center}
\includegraphics[width=4in]{images/select2.pdf}
-\caption{{\bf Selecting multiple columns in an alignment}}
+\caption{{\bf Selecting multiple columns in an alignment.} The red highlighting
+on the alignment ruler marks the selected columns. Note that only the most
+recently selected column has a dashed-box around it to indicate a region
+selection. }
\label{selectcols}
\end{center}
\end{figure}
\begin{figure}[htb]
\begin{center}
\includegraphics[width=4in]{images/select3.pdf}
-\caption{{\bf Selecting multiple sequences in an alignment}}
+\caption{{\bf Selecting multiple sequences in an alignment.} Use [CTRL] or [SHIFT] to select many sequences at once.}
\label{selectrows}
\end{center}
\end{figure}
-\rTo select multiple complete sequences, click and drag the mouse along the sequence ID panel. Multiple sequences will be selected. The same technique as used for columns above can be used with SHIFT-Click and CTRL-Click to select discontinuous ranges of sequences (Figure \ref{selectrows}).\r%[fig 14]
+\rTo select multiple complete sequences, click and drag the mouse down the\rsequence ID panel. The same technique as used for columns above can be used with [SHIFT]-Click for continuous and [CTRL]-Click to select discontinuous ranges of sequences (Figure \ref{selectrows}).\r%[fig 14]
\subsubsection{Making selections in Cursor mode}
-To define a selection in cursor mode, navigate to the top left corner of the proposed selection. Pressing the [Q] key marks this as the corner. A red outline appears around the cursor (Figure \ref{cselect})
+To define a selection in cursor mode (which is enabled by pressing [F2]),
+navigate to the top left corner of the proposed selection (using keystroke
+commands, the arrow keys or the mouse). Pressing the [Q] key marks this as the
+corner. A red outline appears around the cursor (Figure \ref{cselect})
Navigate to the bottom right corner of the proposed selection and press the [M] key. This marks the bottom right corner of the selection. The selection can then be treated in the same way as if it had been created in normal mode.
\subsubsection{Inverting the current selection}
-The current sequence or column selection can be inverted, using {\tr Select $\Rightarrow$ Invert {\bf Sequence/Column} Selection} in the Alignment window. Inverting the selection is particularly useful when hiding regions in a large alignment (see section \ref{hidecol} below). Instead of selecting the columns and rows that are to be hidden, simply select the region that is to be kept visible, and then invert the selection.
-\r\r\subsection{Creating groups}\rSelections are lost as soon as a different region is selected. Groups can be created which are labeled regions of the alignment. To create a group, first select the region which is to comprise the group. Then click the right mouse button on the selection to bring up a context menu. Select {\sl Selection $\Rightarrow$ Group $\Rightarrow$ Group} then enter a name for the group in the dialogue box which appears.
+The current sequence or column selection can be inverted, using {\tr Select
+$\Rightarrow$ Invert {\bf Sequence/Column} Selection} in the Alignment window.
+Inverting the selection is particularly useful when hiding regions in a large
+alignment (see Section \ref{hidecol} below). Instead of selecting the columns
+and rows that are to be hidden, simply select the region that is to be kept
+visible, and then invert the selection.\footnote{It is also possible to hide
+everything but the selected region using the {\sl View $\Rightarrow$ Hide
+$\Rightarrow$ All but selected region } menu entry.}
+\r\r\subsection{Creating groups}\rSelections are lost as soon as a different region is selected. Groups can be
+created which are labeled regions of the alignment. To create a group, first
+select the region which is to comprise the group. Then click the right mouse
+button on the selection to bring up a context menu. Select {\sl Selection
+$\Rightarrow$ Group $\Rightarrow$ Edit name and description of
+current group}\footnote{In earlier versions of Jalview, this entry was variously
+`Group', `Edit Group Name', or `JGroupXXXXX' (Where XXXXX was some serial
+number).} then enter a name for the group in the dialogue box which appears.
-\begin{figure}[htbp]
+\begin{figure}
\begin{center}
-\includegraphics[width=3.5in]{images/group1.pdf}
-\includegraphics[width=2in]{images/newgroup.pdf}
+\includegraphics[width=3in]{images/group1.pdf}
+\includegraphics[width=1.5in]{images/newgroup.pdf}
\caption{{\bf Creating a new group from a selection}}
\label{makegroup}
\end{center}
\end{figure}
-By default the new group will have a box drawn around it. The appearance of the group can be changed (see section \ref{colours} below). This group will stay defined even when the selection is removed.
+By default the new group will have a box drawn around it. The appearance of the group can be changed (see Section \ref{colours} below). This group will stay defined even when the selection is removed.
\subsection{Exporting the current selection}
\exercise{Making selections and groups}{
\label{exselect}
-\exstep{Close all windows in Jalview and load the ferrodoxin alignment (PFAM ID PF03460). Choose a residue and place the mouse cursor on it. Click and drag the mouse cursor to create a selection. As you drag, a red box will 'rubber band' out to show the extent of the selection. Release the mouse button and a red box should border the selected region. Now press [ESC] to clear the selection.}
-\exstep{
+\exstep{Close all windows in Jalview and load the ferredoxin alignment (PFAM
+ID PF03460). Choose a residue and place the mouse cursor on it. Click and drag the mouse cursor to create a selection. As you drag, a red box will `rubber band' out to show the extent of the selection. Release the mouse button and a red box should border the selected region. Now press [ESC] to clear the selection.} \exstep{
Select one sequence by clicking on the id panel. Note that the sequence ID takes on a highlighted background and a red box appears around the selected sequence. Now hold down [SHIFT] and click another sequence ID a few positions above or below. Note how the selection expands to include all the sequences between the two positions on which you clicked.
Now hold down [CTRL] and click on several sequences ID's both selected and unselected. Note how unselected IDs are individually added to the selection and previously selected IDs are individually deselected.
}
\exstep{ Repeat the step above but selecting columns by clicking on the ruler bar instead of selecting rows by clicking on the sequence ID.
}
\exstep{Press [F2] to enter Cursor mode. Navigate to column 59, row 1 by pressing {\sl 5 9 , 1 [RETURN]}. Press {\sl Q} to mark this position. Now navigate to column 65, row 8 by pressing {\sl 6 5 , 8 [RETURN]}. Press {\sl M} to complete the selection.}
-\exstep{
-\label{exselectgrpcolour}
-Open the popup menu by right-clicking the selected region with the mouse. Open the {\sl Selection $\Rightarrow$ Group $\Rightarrow$ Group Colour } menu and select 'Percentage Identity'. This will turn the selected region into a group. }
-\exstep{Hold down [CTRL] and use the mouse to select and deselect sequences by clicking on their Sequence ID label. Note how the group expands to include newly selected sequences, and the 'Percentage Identity' colouring changes. }
+\exstep{\label{exselectgrpcolour}Open the popup menu by right-clicking the
+selected region with the mouse. Open the {\sl Selection $\Rightarrow$ Group $\Rightarrow$ Group Colour } menu and select `Percentage Identity'. This will turn the selected region into a group. }
+\exstep{Hold down [CTRL] and use the mouse to select and deselect sequences by clicking on their Sequence ID label. Note how the group expands to include newly selected sequences, and the `Percentage Identity' colouring changes. }
\exstep{
Use the mouse to click and drag the right-hand edge of the selected group. Note again how the group resizes.}
\subsection{Hiding regions}
-
+\label{hidingregions}
It is sometimes convenient to exclude some sequences or residues in the alignment without actually deleting them. Jalview allows sequences or alignment columns within a view to be hidden, and this facility has been used to create several different views of the example alignment in the file that is loaded when Jalview is first started (See Figure \ref{startpage}).
To hide a set of sequences, select them and right-click the mouse on the selected sequence IDs to bring up the context menu. Select {\sl Hide Sequences} and the sequences will be concealed, with a small triangle indicating their position (Figure \ref{hideseq}). To unhide (reveal) the sequences, right click on the triangle and select {\sl Reveal Sequences} from the context menu.
\end{center}
\end{figure}
-
-
-A similar mechanism applies to columns (Figure \ref{hidecol}). Selected columns (indicated by a red marker) can be hidden and revealed in the same way via the context menu (right click) on the ruler bar. The hidden selection is indicated by a blue triangle in the ruler bar.
+A similar mechanism applies to columns (Figure \ref{hidecol}). Selected columns (indicated by a red marker) can be hidden and revealed in the same way via the context menu (right click) on the ruler bar. The hidden column selection is indicated by a blue triangle in the ruler bar.
\begin{figure}[htbp]
\begin{center}
\end{center}
\end{figure}
+It is often easier to select the region that you intend to work with, rather
+than the regions that you want to hide. In this case, use the {\sl View $\Rightarrow$ Hide
+$\Rightarrow$ All but selected region } menu entry, or press [Shift]+[Ctrl]+H
+to hide the unselected region.
+
\subsubsection{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. However, any alignment edits that affect the sequence will also affect the whole sequence group.
\exercise{Hiding and revealing regions}{
\exstep{Close all windows then open the PFAM accession PF03460. Select a contiguous set of sequences by clicking and dragging on the sequence ID panel. Right click on the selected sequence IDs and select {\sl Hide Sequences}.
}
\exstep{Repeat the above but hiding and revealing columns instead of sequences.
}
-\exstep{Select some sequences and pick one to represent the rest. Bring up the sequence ID pop-up menu for that sequence and select the {\sl Represent group with <Sequence ID> } option. Use the pop-up menu again to reveal the hidden sequences that you just picked a representative for.}
+\exstep{Select a region of the alignment, add in some additional columns to the
+selection, and experiment with the `Hide all but selected region' function. }
+\exstep{Select some sequences and pick one to represent the rest. Bring up the sequence ID pop-up menu for that sequence and select the {\sl Represent group with $<$Sequence ID$>$ } option. Use the pop-up menu again to reveal the hidden sequences that you just picked a representative for.}
}
\subsection{Introducing and removing gaps}
-Jalview provides interactive editing, allowing sequences to be dragged to create gaps.
+The alignment view provides an interactive editing interface, allowing gaps to be inserted or deleted to the left of any position in a sequence or sequence group. Alignment editing can only be performed whilst in keyboard editing mode (entered by pressing [F2]) or by clicking and dragging residues with the mouse when [SHIFT] or [CTRL] is held down (which differs from earlier versions of Jalview).
+
+\subsubsection{Locked Editing}
+\label{lockededits}
+The Jalview alignment editing model is different to that used in other alignment
+editors. Because edits are restricted to the insertion and deletion of gaps to the left of a particular sequence position, editing has the effect of shifting the rest of the sequence(s) being edited down or up-stream with respect to the rest of alignment. The {\sl Edit $\Rightarrow$ Pad Gaps} option can be enabled to eliminate `ragged edges' at the end of the alignment, but does not avoid the `knock-on' effect which is sometimes undesirable. However, its effect can be limited by performing the edit within a selected region. In this case, gaps will only be removed or inserted within the selected region. Edits are similarly constrained when they occur adjacent to a hidden column.
\subsubsection{Introducing gaps in a single sequence}
To introduce a gap, place the cursor on the residue to the immediate right of where the gap should appear. Hold down the SHIFT key and the left mouse button, then drag the sequence to the right till the required number of gaps has been inserted.
-One common error is to forget to hold down [SHIFT]. This results in a selection which is one sequence high and one residue long. Gaps cannot be inserted in such a selection. The selection can be cleared and editing enabled by pressing the [SHIFT] key.
-
-\begin{figure}[htb]
-\begin{center}
-\includegraphics[width=3in]{images/edit1.pdf}
-\includegraphics[width=3in]{images/edit2.pdf}
-\caption{{\bf Introducing gaps.} Gaps are introduced as the selected sequence is dragged to the right.}
-\label{gapseq}
-\end{center}
-\end{figure}
+One common error is to forget to hold down [SHIFT]. This results in a selection which is one sequence high and one residue long. Gaps cannot be inserted in such a selection. The selection can be cleared and editing enabled by pressing the [ESC] key.
\subsubsection{Introducing gaps in all sequences of a group}
To insert gaps in all sequences in a selection or group, place the mouse cursor on any residue in the selection or group to the immediate right of the position in which a gap should appear. Hold down the CTRL key and the left mouse button, then drag the sequences to the right until the required number of gaps has appeared.
-\begin{figure}[htb]
-\begin{center}
-\includegraphics[width=3in]{images/edit3.pdf}
-\includegraphics[width=3in]{images/edit4.pdf}
-\caption{{\bf Introducing gaps.} Gaps are introduced as the selected group is dragged to the right.}
-\label{gapgroup}
-\end{center}
-\end{figure}
-
-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).
+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).
\subsubsection{Sliding Sequences}
-Pressing the [$\Leftarrow$] or [$\Rightarrow$] when one or more sequences are selected will ``slide'' the selected sequences to the left or right (respectively).
+Pressing the [$\leftarrow$] or [$\rightarrow$] arrow keys when one or more
+sequences are selected will ``slide'' the 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.
-\subsubsection{Locked Editing}
-\label{lockededits}
-Normally, when a gap is inserted or removed in the alignment, the whole row is affected. However, if the edit is performed within a selected region, gaps will only be removed or inserted within the selected region. Edits are similarly constrained when they occur adjacent to a hidden column.
-
\subsubsection{Undoing edits}
-Jalview supports the undoing of edits via the {\sl Edit $\Rightarrow$ Undo Edit} alignment window menu option. Each editing action is stored and can be reversed in sequence. Colouring of the alignment is not reversible via the {\sl Undo} option.
+Jalview supports the undoing of edits via the {\sl Edit $\Rightarrow$ Undo Edit}
+alignment window menu option, or CTRL-Z. An edit, if undone, may be re-applied
+with {\sl Edit $\Rightarrow$ Redo Edit}, or CTRL-Y. Note, however, that the
+{\sl Undo} function only works for edits to the alignment or sequence ordering.
+Colouring of the alignment, showing and hiding of sequences or modification of
+annotation cannot be undone.
+
% % better idea to introduce hiding sequences, and use the invert selection, hide
% others, to simplify manual alignment construction
\exercise{Editing alignments}{
\label{mousealedit}
-\exstep{ Load the URL
-\textsf{http://www.jalview.org/tutorial/unaligned.fa} which contains part of the
-ferrodoxin alignment from PF03460.
-
- You are going to manually reconstruct the alignment that jalview loads by
- default. Remember to use [CTRL]+Z to undo an edit, or the {\tr File $\LeftArrow$
+% TODO: VERIFY FOR 2.6.1 and 2.7 - NUMBERING/INSTRUCTIONS APPEAR OFF
+{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}.
+}
+ \item{Remember to use [CTRL]+Z to undo an edit, or the {\tr File $\Rightarrow$
Reload } function to revert the alignment back to the original version if you
- want to start again.
+ want to start again.}
+ \item{ If you are using OSX, and a key combination - such as [CTRL]+A - does
+ not work, then try pressing the [CMD] key instead of [CTRL].
}
+\item{}
+
+\exstep{ Load the URL
+\textsf{http://www.jalview.org/tutorial/unaligned.fa} which contains part of the
+ferredoxin alignment from PF03460.
+}
\exstep{ Select the first 7 sequences, and press H to hide them (or right click
on the sequence IDs to open the sequence ID popup menu, and select {\tr Hide
\exstep{ Select FER1\_SPIOL, FER1\_ARATH, FER2\_ARATH, Q93Z60\_ARATH and
O80429\_MAIZE (Hint: press [CTRL]-I to invert the sequence selection and then
-deleset FER1\_MAIZE), and use the $\Rightarrow$ key to slide them to so they
+deselect FER1\_MAIZE), and use the $\Rightarrow$ key to slide them to so they
begin at column 5 of the alignment view.}
\exstep{ Select all the visible
dragging one column to right. Insert another gap at column 47 in all sequences in
the same way.}
-\exstep{ Correct the feredoxin domain alignment for FER1\_SPIOL by
-insert two additional gaps after the gap at column 47: hold [SHIFT] and click and
-drag on the G and move it two columns to the right.}
+\exstep{ Correct the ferredoxin domain alignment for FER1\_SPIOL by
+insert two additional gaps after the gap at column 47: First press [Escape] to
+clear the selection, then hold [SHIFT] and click and drag on the G and move it
+two columns to the right.}
\exstep{ Now complete the
alignment of FER1\_SPIOL with a {\bf locked edit} by pressing [ESC] and select
\exstep{ Hold [SHIFT] and drag the I at column 39 of FER1\_MAIZE 2 columns to the
right. Remove the gap at FER1\_MAIZE column 49 by [SHIFT]+click and drag left by
-one column. Insert three gaps in FER1\_MAIZE at column 47 by holding [SHIFT] and
-click and drag the S in FER1\_MAIZE to the right by three columns. Finally,
+one column. Press [Escape] to clear the selection, and then insert three gaps in
+FER1\_MAIZE at column 47 by holding [SHIFT] and click and drag the S in FER1\_MAIZE to the right by three columns. Finally,
remove the gap in O80429\_MAIZE at column 56 using [SHIFT]-drag to the left on
42R.}
\exstep{ Use the
-{\sl Edit $\Rightarrow$ Undo Edit} and {\sl Edit $\Rightarrow$ Redo Edit} menu option to step backwards and replay the
-edits you have made}
-}
+{\sl Edit $\Rightarrow$ Undo Edit} and {\sl Edit $\Rightarrow$ Redo Edit} menu
+option, or their keyboard shortcuts ([CTRL]+Z and [CTRL]+Y) to step backwards
+and replay the edits you have made} }
+
+\begin{figure}[htb]
+\begin{center}
+\includegraphics[width=3in]{images/edit1.pdf}
+\includegraphics[width=3in]{images/edit2.pdf}
+\caption{{\bf Introducing gaps in a single sequence.} Gaps are introduced as the
+selected sequence is dragged to the right with [SHIFT] pressed.}
+\label{gapseq}
+\end{center}
+\end{figure}
+
+\begin{figure}[htb]
+\begin{center}
+\includegraphics[width=3in]{images/edit3.pdf}
+\includegraphics[width=3in]{images/edit4.pdf}
+\caption{{\bf Introducing gaps in a group.} Gaps are introduced as the selected
+group is dragged to the right with [CTRL] pressed.}
+\label{gapgroup}
+\end{center}
+\end{figure}
+
\subsubsection{Editing in Cursor mode}
-Gaps can be inserted in cursor mode by pressing [SPACE]. Gaps will be inserted at the cursor, pushing 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 be easily inserted when in cursor mode (toggled with [F2]) by pressing [SPACE]. Gaps will be inserted at the cursor, pushing 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]. 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, use [CTRL]-[BACKSPACE] or [SHIFT]-[BACKSPACE] (both keys held down together).
\exercise{Keyboard edits}{
-\exstep{Load the sequence alignment at \textsf{http://www.jalview.org/tutorial/unaligned.fa}, or continue using the edited alignment from exercise \ref{mousealedit}. If you continue from the previous exercise, then first right click on the sequence ID panel and select {\tr Reveal All}.
+\item{This continues on from exercise \ref{mousealedit}, and recreates the final
+part of the example ferredoxin alignment from the unaligned sequences using
+Jalview's keyboard editing mode.}
+
+\exstep{Load the
+sequence alignment at \textsf{http://www.jalview.org/tutorial/unaligned.fa}, or continue using the edited alignment from exercise \ref{mousealedit}. If you continue from the previous exercise, then first right click on the sequence ID panel and select {\tr Reveal All}.
Now, enter cursor mode by pressing [F2]}
+%TODO: BACKSPACE or DELETE WHEN SEQS ARE SELECTED WILL DELETE ALL SEQS JAL-783
\exstep{Insert 58 gaps at the start of the first sequence (FER\_CAPAA). Press {\sl 58} then {\sl [SPACE]}. }
\exstep{Go down one sequence and select rows 2-5 as a block. Click on the second sequence ID (FER\_CAPAN). Hold down shift and click on the fifth (FER1\_PEA). }
\exstep{Insert 6 gaps at the start of this group. Go to column 1 row 2 by typing {\sl 1,2} then pressing {\sl [RETURN]}. Now insert 6 gaps. Type {\sl 6} then hold down [CTRL] and press the space bar.}
\r\section{Colouring sequences}
\label{colours}
-Colouring sequences is a key aspect of alignment presentation. Jalview allows both for colouring of the whole alignment and colouring of selections/groups. There are two main types of colouring styles: simple static residue colourschemes, dynamic schemes which use conservation and consensus analysis to control colouring. A hybrid colouring is also possible, where static residue schemes are modified using a dynamic scheme. The individual schemes are described in section \ref{colscheme} below.
+Colouring sequences is a key aspect of alignment presentation. Jalview allows
+you to colour the whole alignment, or just specific groups. Alignment and
+group colours are rendered
+{\sl below} any other colours, such as those arising from sequence features
+(these are described in Section \ref{featannot}). This means that if you
+try to apply one of the colourschemes described in this section, and nothing
+appears to happen, it may be that you have sequence feature annotation
+displayed, and you may have to disable it using the {\sl View $\Rightarrow$
+Show Features} option before you can see your colourscheme.
+
+There are two main types of colouring styles: simple static residue colourschemes and dynamic schemes which use conservation and consensus analysis to control colouring. A hybrid colouring is also possible, where static residue schemes are modified using a dynamic scheme. The individual schemes are described in Section \ref{colscheme} below.
\subsection{Colouring the whole alignment}
-\parbox[c]{3.75in}{The alignment can be coloured via the {\sl Colour} menu option in the alignment window. Selecting the colour scheme causes all residues to be coloured. The menu is divided into two three sections. The first section gives options for the behaviour of the menu options, the second lists static and dynamic colourschemes available for selection, and the last gives options for making hybrid colourschemes using conservation shading or colourscheme thresholding.
+\parbox[c]{3.75in}{The alignment can be coloured via the {\sl Colour} menu option in the alignment window. Selecting the colour scheme causes all residues to be coloured. The menu is divided into three sections. The first section gives options for the behaviour of the menu options, the second lists static and dynamic colourschemes available for selection. The last gives options for making hybrid colourschemes using conservation shading or colourscheme thresholding.
}\parbox[c]{3in}{
\centerline {
\subsection{Colouring a group or selection}
-Selections or groups can be coloured via two ways. The first is via the alignment Colour menu but ensuring that the Apply to all groups flag is not selected. This will have to be turned off specifically as it is on by default.
+Selections or groups can be coloured in two ways. The first is via the Alignment Window's {\sl Colour} menu, after first ensuring that the Apply to all groups flag is not selected. This must be turned off specifically as it is on by default.
The second method is to use the {\sl Selection $\Rightarrow$ Group $\Rightarrow$ Group Colour} context menu option obtained by right clicking on the group (Figure \ref{colgrp}).
\begin{figure}[htbp]
\begin{center}
+%TODO update group_col.pdf to show latest jalview group edit submenu
\includegraphics[width=4in]{images/group_col.pdf}
\caption{{\bf Colouring a group via the context menu.}}
\label{colgrp}
\end{figure}
\subsection{Shading by conservation}
-For many colour schemes, the intensity of the colour in a column can be scaled by the degree of amino acid property conservation. Selecting {\sl Colour $\Rightarrow$ By Conservation} brings up a selection box (the {\sl Conservation Threshold dialog box}) allowing the alignment colouring to be modified. Selecting a higher value limits colouring to higher conserved groups (Figure \ref{colcons}).
+For many colour schemes, the intensity of the colour in a column can be scaled
+by the degree of amino acid property conservation. Selecting {\sl Colour
+$\Rightarrow$ By Conservation} enables this mode, and {\sl Modify
+conservation threshold} brings up a selection box (the {\sl Conservation
+Threshold dialog box}) allowing the alignment colouring to be modified.
+Selecting a higher value limits colouring to more highly conserved columns (Figure \ref{colcons}).
\begin{figure}[htbp]
\begin{center}
\subsection{Thresholding by percentage identity}
-'Thresholding' is another hybrid colour model where a residue is only coloured if it is not excluded by an applied threshold. Selecting {\sl Colour $\Rightarrow$ Above Identity Threshold} brings up a selection box with a slider controlling the minimum percentage identity threshold to be applied. Selecting a higher threshold (by sliding to the right) limits the colouring to columns with a higher percentage identity (as shown by the Consensus histogram in the annotation panel).
+`Thresholding' is another hybrid colour model where a residue is only coloured
+if it is not excluded by an applied threshold. Selecting {\sl Colour $\Rightarrow$ Above Identity Threshold} brings up a selection box with a slider controlling the minimum percentage identity threshold to be applied. Selecting a higher threshold (by sliding to the right) limits the colouring to columns with a higher percentage identity (as shown by the Consensus histogram in the annotation panel).
\subsection{Colouring by Annotation}
+\label{colourbyannotation}
\parbox[c]{3in}{
-Any of the quantitative annotation on an alignment can be used to threshold or shade the whole alignment. The {\sl Colour $\Rightarrow$ By Annotation} options opens a dialog which allows you to select which annotation to use, the minimum and maximum shading colours or whether an existing colouring is to be thresholded.
-}\parbox[c]{3in}{
+Any of the quantitative annotations shown on an alignment can be used to
+threshold or shade the whole alignment\footnote{Please remember to turn off
+Sequence Feature display to see the shading}. The {\sl Colour $\Rightarrow$ By
+Annotation} options opens a dialog which allows you to select which annotation
+to use, the minimum and maximum shading colours or whether the original
+colouring should be thresholded (the `Use original colours' option).
+
+Default settings for the minimum and maximum colours can be set in the Jalview
+Desktop's preferences. }\parbox[c]{3in}{
\centerline{\includegraphics[width=2.5in]{images/col_byannot.pdf}}}
\subsection{Colour schemes}
\subsubsection{Helix Propensity}
\parbox[c]{3.5in}{
-The residues are coloured according to their helix propensity. The highest propensity is magenta, the lowest is green.
+The residues are coloured according to their Chou-Fasman\footnote{\label{chou-fasman}Chou, PY and Fasman, GD. Annu Rev Biochem. 1978;47:251-76.} helix propensity. The highest propensity is magenta, the lowest is green.
}
\parbox[c]{3in}{
\includegraphics[width=2.75in]{images/col_helix.pdf}
\subsubsection{Strand Propensity}
\parbox[c]{3.5in}{
-The residues are coloured according to their Strand propensity. The highest propensity is Yellow, the lowest is blue.
+The residues are coloured according to their Chou-Fasman\textsuperscript{\ref{chou-fasman}} Strand propensity. The highest propensity is Yellow, the lowest is blue.
}
\parbox[c]{3in}{
\includegraphics[width=2.75in]{images/col_strand.pdf}
\subsubsection{Turn Propensity}
\parbox[c]{3.5in}{
-The residues are coloured according to their turn propensity. The highest propensity is red, the lowest is cyan.
+The residues are coloured according to their Chou-Fasman\textsuperscript{\ref{chou-fasman}} turn propensity. The highest propensity is red, the lowest is cyan.
}
\parbox[c]{3in}{
\includegraphics[width=2.75in]{images/col_turn.pdf}
\subsubsection{Buried Index}
\parbox[c]{3.5in}{
-The residues are coloured according to their burial propensity. The highest propensity is blue, the lowest is green.
+The residues are coloured according to their Chou-Fasman\textsuperscript{\ref{chou-fasman}} burial propensity. The highest propensity is blue, the lowest is green.
}
\parbox[c]{3in}{
\includegraphics[width=2.75in]{images/col_buried.pdf}
\subsubsection{Nucleotide}
\parbox[c]{3.5in}{
-Residues are coloured with four colours corresponding to the four nucleotide bases. All non ACTG residues are uncoloured. See section \ref{workingwithnuc} for further information about working with nucleic acid sequences and alignments.
+Residues are coloured with four colours corresponding to the four nucleotide bases. All non ACTG residues are uncoloured. See Section \ref{workingwithnuc} for further information about working with nucleic acid sequences and alignments.
}
\parbox[c]{3in}{
\includegraphics[width=2.75in]{images/col_nuc.pdf}
}
}
\r\section{Alignment formatting and graphics output}
-
+\label{layoutandoutput}
Jalview is a WYSIWIG alignment editor. This means that for most kinds of graphics output, the layout that is seen on screen will be the same as what is output in an exported graphics file. It is therefore important to pick the right kind of display layout prior to generating figures.
\subsection{Multiple Alignment Views}
Jalview is able to create multiple independent visualizations of the same underlying alignment - these are called {\tr 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 {\tr View $\RightArrow$ New View} option of the alignment window. This will create a new view with the same groups, alignment layout and display options as the current one. Views may be gathered (by pressing G) together as named tabs on the alignment window, or displayed simultaneously in their own window (by pressing X).}\parbox[c]{2.75in}{
+\parbox[c]{4in}{Alignment views are created using the {\tr View $\Rightarrow$ New View} option of the alignment window. This will create a new view with the same groups, alignment layout and display options as the current one. Views may be gathered (by pressing G) together as named tabs on the alignment window, or displayed simultaneously in their own window (by pressing X).}\parbox[c]{2.75in}{
\begin{center}\centerline{
\includegraphics[width=2.5in]{images/mulv_tabs.pdf}}
\end{center}
% JBPNote make an excercise on views ?
\subsection{Alignment layout}
-Jalview provides two screen layout modes, unwrapped (the default) where the alignment is in one long line across the window, and wrapped, where the alignment is on multiple lines, each the width of the window. Most layout options are controlled by the Format menu option in the alignment window.
+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. Furthermore, alignment annotation (see section \ref{featannot}) cannot be interactively created or edited in wrapped mode, and selection of large regions is difficult.
+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. Furthermore, alignment annotation (see Section \ref{featannot}) cannot be interactively created or edited in wrapped mode, and selection of large regions is difficult.
\begin{figure}[htbp]
\begin{center}
-% this view still refers to Jalview 2.2 and the menu is missing the Center Columns option
\parbox[c]{2in}{\includegraphics[width=2in]{images/wrap1.pdf}}
\parbox[c]{4in}{\includegraphics[width=4in]{images/wrap2.pdf}}
\caption{{\bf Wrapping the alignment}}
\subsubsection{Fonts}
-\parbox[c]{4in}{Text appearance can be modified via the {\sl Format $\Rightarrow$ Font\ldots} alignment window menu. Additionally, font size and spacing can be adjusted rapidly by clicking the middle mouse button and dragging across the alignment window.}
+\parbox[c]{4in}{The text appearance in a view can be modified via the {\sl Format $\Rightarrow$ Font\ldots} alignment window menu. This setting applies for all alignment and annotation text except for that displayed in tool-tips. Additionally, font size and spacing can be adjusted rapidly by clicking the middle mouse button and dragging across the alignment window.}
\parbox[c]{2in}{\centerline{\includegraphics[width=1.75in]{images/font.pdf}}}
-\subsubsection{Numbering and annotations}
-
-The {\sl Format} menu also provides options to control the display of numbers and annotations on sequence alignments.
-The annotation lines which appear below the sequence alignment can be hidden by toggling the {\sl View $\Rightarrow$ Show Annotations} menu option. Additionally, each annotation line can be hidden and revealed in the same way as sequences via the context menu on the annotation name panel (Figure \ref{annot}). Annotations can be reordered by dragging the annotation line label on the annotation label panel. Placing the mouse over the top annotation label brings up a resize icon. Dragging the top annotation line label up and down alters the relative size of the sequence alignment and annotation alignment panels.
+\subsubsection{Numbering and label justification}
+Options in the {\sl Format} menu are provided to control the also provides a range of options to control the display of sequence and alignment numbering, the justification of sequence IDs and annotation row column labels on the annotation rows shown below the alignment.
+
+\subsubsection{Alignment and Group colouring and appearance}
+The display of hidden row/column markers and gap characters can be turned off with {\sl Format $\Rightarrow$ Hidden Markers} and {\sl Format $\Rightarrow$ Show Gaps}, respectively. The {\sl Text} and {\sl Colour Text} option controls the display of sequence text and the application of alignment and group colouring to it. {\sl Boxes } controls the display of the background area behind each residue that is coloured by the applied coloursheme.
+
+\subsubsection{Highlighting nonconserved symbols}
+The alignment layout and group sub-menu both contain an option to hide
+conserved symbols from the alignment display ({\sl Format $\Rightarrow$ Show
+nonconserved } in the alignment window or {\sl Popup Menu $\Rightarrow$ Group
+$\Rightarrow$ Show nonconserved}). This mode is useful when working with
+alignments that exhibit a high degree of homology, because Jalview will only
+display gaps or sequence symbols that differ from the consensus for each
+column, and render all others with a `.'.
+%TODO add a graphic to illustrate this.
+\subsection{Annotation ordering and display}
+% TODO: describe consensus, conservation, quality user preferences, and group
+% annotation preferences.
+The annotation lines which appear below the sequence alignment are described in
+detail in Section \ref{annot}. They can be hidden by toggling the {\sl View
+$\Rightarrow$ Show Annotations} menu option. Additionally, each annotation line
+can be hidden and revealed in the same way as sequences via the context menu on
+the annotation name panel (Figure \ref{annot}). Annotations can be reordered by
+dragging the annotation line label on the annotation label panel. Placing the
+mouse over the top annotation label brings up a resize icon. When this is
+displayed, Click-dragging up and down alters the relative size of the sequence
+alignment and annotation alignment panels.
\begin{figure}
\begin{center}
\exstep{Start Jalview and open the URL \textsf{http://www.jalview.org/examples/exampleFile.jar}. Select {\sl Format $\Rightarrow$ Wrap} from the alignment window menu. Experiment with the various options from the {\sl Format} menu. to adjust the ruler placement, sequence ID format and so on. }
\exstep{Hide all the annotation rows by selecting {\sl View $\Rightarrow$ Show Annotations} from the alignment window menu. Reveal the annotations by selecting the same menu option.}
\exstep{Right click on the annotation row labels to bring up the context menu. 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} below.}
+\exstep{Annotations can be reordered by clicking and dragging the row to the desired position. Click on the {\sl Consensus} row and drag it upwards to just above {\sl Quality}. The rows should now be reordered. Features and annotations are covered in more detail in Section \ref{featannot} below.}
\exstep{Move the mouse to the top left hand corner of the Secondary Structure annotation row label - a grey up/down arrow symbol should appear - when this is shown, the height of the {\sl Annotation Area} can be changed by Clicking and dragging the mouse up or down.}
}
\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 via the {\sl File $\Rightarrow$ Export Image $\Rightarrow$ \ldots } alignment window menu option. }
\parbox[c]{2in}{\centerline{\includegraphics[width=1.25in]{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 seuqence'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]{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]{3.5in}{\centerline{\includegraphics[width=3in]{images/image_html.pdf}}}
\subsubsection{EPS}
-\parbox[c]{3in}{EPS is Encapsulated Postscript. It is the format of choice for publication and posters as it gives the highest quality output of any of the image types. It can be scaled indefinitely 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.
+\parbox[c]{3in}{EPS is Encapsulated Postscript. It is the format of choice for publication and posters as it gives the highest quality output of any of the image types. It can be scaled indefinitely 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.
}
\parbox[c]{3.5in}{\centerline{\includegraphics[width=3in]{images/image_eps.pdf}} \par \centerline{Zoom Detail of EPS image.}}
\subsubsection{PNG}
\parbox[c]{3in}{
-PNG is Portable Network Graphics. This output option produces an image that can be easily included in web pages and incorporated in presentations using e.g. Powerpoint or Open Office. It is a bitmap image so does not scale and is unsuitable for use on posters.
+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.}}
-
-\exercise{Graphical Output}{
+ \exercise{Graphical Output}{
\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. Save the file and open it in your favourite web browser. }
\exstep{Now wrap the alignment (Exercise \ref{exscreen}) and export the image to HTML again. Compare the two images. Note that the exported image matches the format displayed in the alignment window but annotations are not exported.}
\exstep{Export the alignment using the {\sl File $\Rightarrow$ Export Image $\Rightarrow$ PNG} menu option. Open the file in an image viewer that allows zooming (eg. Paint or Photoshop on Windows, Preview on Mac OS X) and zoom in. Notice that the image is a bitmap and it becomes pixelated very quickly. Note also that the annotation lines are included in the image.}
}
\chapter{Analysis and Annotation}
+\label{analysisannotation}
-\section{Analysis of alignments}
-Jalview provides support for sequence analysis in two ways. A number of analytical methods are 'built-in' and run inside Jalview itself and are mostly accessed from the {\sl Calculate} alignment window menu. Computationally intensive analyses are run outside Jalview via web services - these are typically accessed via the {\sl Web Services} menu.
+This chapter describes the annotation, analysis, and visualization tasks that the Jalview Desktop can perform. Section \ref{wkwithstructure} introduces the structure visualization capabilities of Jalview. In Section \ref{alignanalysis}, you will find details of the Tree building, viewing and PCA capabilities, alignment redundancy removal, pairwise alignments and alignment conservation analysis. Subsequently, in Section \ref{jvwebservices}, programs available remotely for multiple sequence alignment and secondary structure prediction are described.
+
+Section \ref{featannot} describes the mechanisms provided by Jalview for interactive creation of sequence and alignment annotation and how they are displayed, imported and exported. Section \ref{featuresfromdb} discusses the retrieval of database references and establishment of sequence coordinate systems for the retrieval and display of features from databases and DAS annotation services. Finally, Section \ref{workingwithnuc} describes functions and visualization techniques relevant to working with nucleotide sequences, coding region annotation and nucleotide sequence alignments.
+
+\section{Working with structures}
+\label{wkwithstructure}
+Jalview facilitates the use of protein structures for the analysis of alignments by providing a linked view of structures associated with protein sequences in the alignment. The Java based molecular viewing program Jmol\footnote{See the Jmol homepage \url{http://www.jmol.org} for more information.} has been incorporated\footnote{Earlier versions of Jalview included MCView - a simple main chain structure viewer. Structures are visualized as an alpha carbon trace and can be viewed, rotated and coloured in a structure viewer and the results interpreted on a sequence alignment.} which enables sophisticated molecular visualizations to be prepared and investigated alongside an analysis of associated sequences.
+PDB format files can be imported directly or structures can be retrieved from
+the European Protein Databank (PDBe) using the Sequence Fetcher (see
+\ref{fetchseq}).
+
+\subsection{Automatic association of PDB structures with sequences}
+Jalview can automatically determine which structures are associated with a
+sequence in a number of ways.
+\subsubsection{Discovery of PDB ids from sequence database cross-references}
+if a sequence has an ID from a public database that contains cross-references to
+the PDB, such as Uniprot. Right-click on any sequence ID and select {\sl $<$Sequence ID$>$ $\Rightarrow$ Structure $\Rightarrow$
+Associate Structure with Sequence $\Rightarrow$ Discover PDB IDs } from the context menu (where {\sl $<$Sequence ID$>$} is the ID of the sequence on which
+you clicked) (Figure \ref{auto}). Jalview will attempt to associate the
+sequence with a Uniprot sequence and from there discover any associated PDB
+structures. This takes a few seconds and applies to all sequences in the
+alignment which have valid Uniprot IDs. On moving the cursor over the sequence
+ID the tool tip\footnote{Tip: The sequence ID tooltip can often become large for
+heavily cross referenced sequence IDs. Use the {\sl View $\Rightarow$ 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.
+
+\begin{figure}[htbp]
+\begin{center}
+%TODO fix formatting
+\parbox{1.5in}{
+{\centering
+\begin{center}
+\includegraphics[width=1.5in]{images/auto1.pdf}
+\end{center}}
+} \parbox{3.25in}{
+{\centering
+\begin{center}
+\includegraphics[scale=0.5]{images/auto2.pdf}
+\end{center}
+}
+} \parbox{1.5in}{
+{\centering
+\begin{center}
+\includegraphics[width=1.5in]{images/auto3.pdf}
+\end{center}
+}
+}
+
+\caption{{\bf Automatic PDB ID discovery.} The tooltip (left) indicates that no PDB structure has been associated with the sequence. After PDB ID discovery (center) the tool tip now indicates the Uniprot ID and any associated PDB structures (right)}
+\label{auto}
+\end{center}
+\end{figure}
+
+\subsubsection{Drag-and-drop association of PDB files with sequences by filename
+match}
+\label{multipdbfileassoc}
+If one or more PDB files are dragged from a file browser onto an alignment
+window, Jalview will search the alignment for sequences with IDs that
+matches 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 no associations are made, then sequences extracted
+from the structure will be simply appended to the alignment. However, if only
+some of the files are associated, jalview will raise another dialog box giving
+you the option to add any remaining sequences the structure files not present in
+the alignment. This allows you to easily decorate sequences in a newly imported
+alignment with any corresponding structures you've collected in a directory
+accessible from your computer.\footnote{We plan to extend this facility in
+future so Jalview will automatically search for PDB files matching your
+sequence within a local directory. Check out
+\href{http://issues.jalview.org/browse/JAL-801}{Jalview issue 801}}
-This section describes the built in analyses. Web Services are described in a subsequent section.
+\begin{figure}[htbp]
+\begin{center}
+\includegraphics[]{images/pdbdragdropassoc.pdf}
+
+\caption{{\bf Associating PDB files with sequences by drag-and-drop.} Dragging
+PDB files to an alignment of sequences with names matching the dragged
+files names (A), results in a dialog box (B) that gives the option to associate each file with any sequences with matching IDs. }
+\label{multipdbfileassocfig}
+\end{center}
+\end{figure}
+
+
+\subsection{Viewing Protein Structures}
+The structure viewer can be launched in two ways from the sequence ID context
+menu. To view a particular structure associated with a sequence in the
+alignment, simply select it from popup menu's associated structures submenu in
+{\sl Structure $\Rightarrow$ View Structure $\Rightarrow$ $<$PDB ID$>$}. The
+second way is most useful if you want to view all structural data available for
+a set of sequences in an alignment. If any of the {\bold currently selected}
+sequences have structures associated, the {\sl Structure } submenu of the
+sequence ID popup menu will include an option to {\sl View {\bold N}
+structures}. Selecting this option will open a new structure view containing
+the associated structures superposed according to the alignment.
+
+In both cases, each structure to be displayed will be downloaded or loaded from
+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 (if available).
+Moving the mouse cursor over a sequence to which the structure is linked in the
+alignment panel highlights the respective residue's sidechain atoms. The
+sidechain highlight may be obscured by other parts of the molecule. Similarly,
+moving the cursor over the structure shows a tooltip and highlights the
+corresponding residue in the alignment. Often, the position highlighted may not
+be in the visible portion of the current alignment view. If the alignment
+window's {\sl View $\Rightarrow$ Automatic Scrolling } option is not selected,
+then you may have to manually move the alignment scroll bars to see the
+highlighted region.
+
+\begin{figure}[htbp]
+\begin{center}
+\parbox{3in}{
+{\centering
+\begin{center}
+\includegraphics[scale=0.5]{images/structure1.pdf}
+\end{center}
+}
+}
+\parbox{3.2in}{
+{\centering
+\begin{center}
+\includegraphics[width=3in]{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). }
+\label{structure}
+\end{center}
+\end{figure}
+
+\subsection{Customising structure display}
+
+Structure display can be modified using the {\sl Colour} and {\sl View} menus
+in the structure viewer. The background colour can be modified by selecting the
+{\sl Colours $\Rightarrow$ Background Colour\ldots} option.
+
+By default, the structure will be coloured in the same way as the
+associated sequence(s) in the alignment view from which it was launched. The
+structure can be coloured independently of the sequence by selecting an
+appropriate colour scheme from the {\sl Colours} menu. It can be coloured
+according to the alignment using the {\sl Colours $\Rightarrow$ By Sequence } option. The image in the structure viewer can be
+output to EPS or PNG format via the {\sl File $\Rightarrow$ Save As
+$\Rightarrow$ \ldots} submenu. The mapping between the structure and the
+sequence (How well and which parts of the structure relate to the sequence)
+can be viewed with the {\sl File $\Rightarrow$ View Mapping} menu option.
+
+\subsubsection{Using the Jmol visualization interface }
+
+Jmol has a comprehensive set of selection and visualization functions that are
+accessed from the Jmol popup menu (by right-clicking in the Jmol window or by
+clicking the Jmol logo). Molecule colour and rendering style can be
+manipulated, and distance measurements and molecular surfaces can be added to
+the view. It also has its own ``Rasmol\footnote{see
+http://www.rasmol.org}-like'' scripting language, which is described elsewhere\footnote{Jmol Wiki: \url{http://wiki.jmol.org/index.php/Scripting}
+
+Jmol Scripting reference:
+\url{http://www.stolaf.edu/academics/chemapps/jmol/docs/}}. Jalview utilises the
+scripting language to interact with Jmol and to store the state of a Jmol visualization within Jalview archives, in addition to the PDB data file originally loaded or retrieved by Jalview. To access the Jmol scripting environment directly, use the {\sl Jmol $\Rightarrow$ Console} menu option.
+
+If you would prefer to use Jmol to manage structure colours, then select the
+{\sl Colours $\Rightarrow$ Colour with Jmol} option. This will disable any
+automatic application of colour schemes when new structure data is added, or
+when associated alignment views are modified.
+
+
+\exercise{Viewing Structures}{\label{viewingstructex}
+\exstep{Load the alignment at
+\textsf{http://www.jalview.org/examples/exampleFile.jar}. Right-click on the
+sequence ID label for any of the sequences (e.g. {\sl FER1\_SPIOL}) to bring up
+the context menu. Select {\sl FER1\_SPIOL $\Rightarrow$ Structure $\Rightarrow$
+Associate Structure with Sequence $\Rightarrow$ Discover
+PDB ids}. Jalview will now attempt to find PDB structures for the sequences in
+the alignment. } \exstep{ Right-click on the sequence ID for {\sl FER1\_SPIOL}.
+Select { \sl FER1\_SPIOL $\Rightarrow$ Structure $\Rightarrow$ View Structure
+$\Rightarrow$ 1A70} A structure viewing window appears. Rotate the molecule by clicking and dragging in the structure viewing box. Zoom with the mouse scroll wheel. } \exstep{Roll the mouse cursor along the {\sl FER1\_SPIOL} sequence in the alignment. Note that if a residue in the sequence maps to one in the structure, a label will appear next to that residue in the structure viewer. Move the mouse over the structure. Placing the mouse over a part of the structure will bring up a tool tip indicating the name and number of that residue. The corresponding residue in the sequence is highlighted in black. Clicking the alpha carbon toggles the highlight and residue label on and off. Try this by clicking on a set of three or four adjacent residues so that the labels are persistent, then finding where they are in the sequence. } \exstep{Select {\sl Colours $\Rightarrow$ Background Colour\ldots} from the structure viewer menu and choose a suitable colour. Press {\sl OK} to apply this. Select {\sl File $\Rightarrow$ Save As $\Rightarrow$ PNG} and save the image. View this with your web browser. }
+\exstep{Select {\sl File $\Rightarrow$ View Mapping} from the structure viewer menu. A new window opens showing the residue by residue alignment between the sequence and the structure.}
+
+\exstep{Select {\sl File $\Rightarrow$ Save $\Rightarrow$ PDB file} and choose a new filename to save the PDB file. Once the file is saved, open the location in your file browser (or explorer window) and drag the PDB file that you just saved on to the Jalview desktop (or load it from the {\sl Jalview Desktop $\Rightarrow$ Input Alignment $\Rightarrow$ From File } menu). Verify that you can open and view the associated structure from the sequence ID pop-up menu's {\sl Structure } submenu in the new alignment window.}
+
+\exstep{Right click on the structure to bring up the Jmol window. Explore the menu options. Try to change the style of molecular display - by first using the {\sl Jmol $\Rightarrow$ Select $\Rightarrow$ all} command, and then the {\sl Jmol $\Rightarrow$ Style $\Rightarrow$ Scheme $\Rightarrow$ Ball and stick} command.}
+\exstep{Use the {\sl File $\Rightarrow$ Save As .. } function to save the alignment as a Jalview Project. Now close the alignment and the structure view, and load the project file you just saved.
+
+Verify that the Jmol display is as it was when you just saved the file.}
+}
+
+\subsection{Superimposing structures}
+\label{superposestructs}
+Many comparative biomolecular analysis investigations aim to determine if the
+biochemical properties of a given molecule are signficantly different to its
+homologues. When structure data is available, comparing the shapes of molecules
+by superimposing them enables substructure that may impart different behaviour
+to be quickly identified. The identification of optimal 3D superpositions
+involves aligning 3D data rather than sequence symbols, but the result can
+still be represented as a sequence alignment, where columns indicate positions
+in each molecule that should be superposed to recreate the optimal 3D alignment.
+
+Jalview can employ Jmol's 3D fitting routines\footnote{See
+\href{http://chemapps.stolaf.edu/jmol/docs/?ver=12.2#compare}{http://chemapps.stolaf.edu/jmol/docs/?ver=12.2$\#$compare}
+for more information.} to recreate 3D structure superpositions based on the
+correspondences defined by one or more sequence alignments involving structures shown in the Jmol display. Superposition based on the currently displayed alignment view happens automatically if a
+structure is added to an existing Jmol display using the {\sl Structure
+$\Rightarrow$ View PDB Structure $\Rightarrow$ ..}. A new Jmol view containing
+superposed structures can also be created using the {\sl Structure
+$\Rightarrow$ View all {\bf N} PDB Structures} option (when {\bf {\sl N}}
+$>$ 1) if the current selection contains two or more sequences with associated
+structures.
+
+\subsubsection{Obtaining the RMSD for a superposition}
+Figure \ref{mstrucsuperposition} shows a superposition created during the course
+of Exercise \ref{superpositionex}. The parts of each molecule used to construct
+the superposition are rendered using the cartoon style, with other parts of
+the molecule drawn in wireframe. The Jmol console, which has been opened after
+the superposition was performed, shows the RMSD report for the superposition.
+Full information about the superposition is also output to the Jalview console,
+including the precise atom pairs used to superpose structures.
+
+\subsubsection{Choosing which part of the alignment is used for structural
+superposition}
+Jalview uses the visible part of each alignment view to define which parts of
+each molecule are to be corresponded. Hiding a column in a view used for
+superposition will remove that correspondence from the set, allowing
+the selection of specific parts of the alignment to be used for
+superposition. Only columns that define a complete set of correspondences for
+all structures will be used for structural superposition, and as a consequence,
+the RMSD values generated for each pair of structures superimposed can be
+directly compared.
+
+In order to recompute a superposition after changing a view or editing the
+alignment, select the {\sl Jmol $\Rightarrow$ Align } menu option. The {\sl
+Jmol $\Rightarrow$ Superpose with ..} submenu allows you to choose which of the
+associated alignments and views are to be used to create the set of
+correspondences. This menu is useful when composing complex superpositions
+involving multi-domain and multi-chain complexes, when correspondences may be
+defined by more than one alignment.
+
+
+\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}
+
+\exercise{Aligning structures using the ferredoxin
+sequence alignment.}{\label{superpositionex}
+
+\exstep{Continue with the Jalview project created in exercise
+\ref{viewingstructex}. Use the {\sl Discover PDB IDs} function to retrieve PDB
+IDs associated with the FER1\_MAIZE sequence.}
+\exstep{Once discovery has completed, use the {\sl
+View PDB Structure} submenu to view the PDB file associated with FER1\_MAIZE.
+Jalview will give you the option of aligning the structure to the one already
+open. To superimpose the structure associated with FER1\_MAIZE with the one
+associated with FER1\_SPIOL, press the {\bf Yes} button.
+
+{\sl The Jmol view will update to show both structures, and one will be
+moved on to the other.}}
+\exstep{Create a new view on the alignment, and hide all but columns 121
+through to 132.}
+\exstep{Use the {\sl Jmol} submenu to
+recompute the superposition using just columns 121-132 of the alignment.
+
+{\sl Note how the molecules shift position when superposed using a short part of
+the two structures.}}
+\exstep{Compare the initial and final RMSDs for superimposing molecules with
+the small section and with the whole alignment. Which view do you think give the
+best 3D superposition, and why ?} }
+
+\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
+sequences associated with structure data shown in a Jmol have multiple views, Jalview gives you full control
+over which alignment, or alignment view, is used to colour the structure
+display. Sequence-structure colouring associations are
+changed {\sl via} the {\sl View $\Rightarrow$ Colour by} menu, which lists all
+views associated with data shown in the embedded Jmol view. A tick is shown beside
+views currently used as colouring source, and moving the
+mouse over each view will bring it to the front of the alignment display,
+allowing you to browse available colour sources prior to selecting one. If the
+{\sl Select many views} option is selected, then multiple views can be selected as sources for colouring the
+structure data. {\sl Invert selection} and {\sl Select all} options are also provided to quickly change between multi-view selections.
+
+\begin{figure}[htbp]
+\begin{center}
+\includegraphics[width=5.5in]{images/mviewstructurecol.pdf}
+\caption{{\bf Choosing a different view for colouring a structure display}
+Browsing the {\sl View $\Rightarrow$ Colour by ..} menu provides full control
+of which alignment view is used to colour structures when the {\sl Colour
+$\Rightarrow$ Colour by sequence} option is selected.}
+\label{mviewstructurecol}
+\end{center}
+\end{figure}
+
+\subsubsection{Colouring complexes}
+
+The ability to control which multiple alignment or view is used to colour
+structural data is essential when working with data relating to
+multidomain biomolecules and complexes.
+
+In these situations, each chain identified in the structure may have a different
+evolutionary history, and a complete picture of functional variation can
+only be gained by integrating data from different alignments on the same
+structure view. An example of this is shown in Figure
+\ref{mviewalcomplex}, based on data from Song et. al\footnote{Structure of
+DNMT1-DNA Complex Reveals a Role for Autoinhibition in Maintenance DNA Methylation. Jikui Song, Olga Rechkoblit, Timothy H. Bestor, and Dinshaw J. Patel.
+{\sl Science} {\bf 331} 1036-1040
+\href{http://www.sciencemag.org/content/331/6020/1036}{DOI:10.1126/science.1195380}}
+
+\begin{figure}[htbp]
+\begin{center}
+\includegraphics[]{images/mchainstructureview.pdf}
+\caption{{\bf The biological assembly of Mouse DNA Methyltransferase-1 coloured
+by Pfam alignments for its major domains} Alignments for each domain within the
+Uniprot sequence DNMT1\_MOUSE have been used to visualise sequence conservation
+in each component of this protein-DNA complex. Instructions for recreating this figure are given in exercise \ref{dnmtcomplexex}. }
+\label{mviewalcomplex}
+\end{center}
+\end{figure}
+
+\exercise{Colouring a protein complex to explore domain-domain interfaces}{\label{dnmtcomplexex}
+
+\exstep{Download the PDB file at
+\textsf{http://www.jalview.org/tutorial/DNMT1\_MOUSE.pdb} to your desktop. This
+is the biological unit for PDB ID 3pt6, as identified by the PDBe's PISA server.}
+
+\exstep{Launch the Jalview desktop and ensure you have at least 256MB of
+free memory available.
+
+{\sl Use the following webstart link:
+\href{http://www.jalview.org/webstart/jalview_1G.jnlp}{http://www.jalview.org/webstart/jalview\_1G.jnlp}}.}
+\exstep{Retrieve the following
+{\bf full} PFAM alignments: PF02008, PF00145, PF01426. These will each be retrieved into their own alignment window.}
+\exstep{Drag the structure you downloaded in
+step 1 onto one of the alignments to associate it with the mouse sequence in
+that Pfam domain family.}
+\exstep{For every DNMT1\_MOUSE sequence in the alignment, use the sequence
+ID popup menu's structure submenu to view the DNMT1\_MOUSE structure for the associated mouse sequence. Check the contents of
+the {\sl View $\Rightarrow$ Colour by ..} submenu to see what alignments can be used to
+colour the sequence.}
+\exstep{Repeat the previous two steps for each of
+the other alignments. In each case, when performing the `View DNMT1\_MOUSE.pdb'
+step, Jalview will ask you if you wish to create a new Jmol view. You should
+respond `No', ensuring that each sequence fragment is associated with the same Jmol view.}
+\exstep{Pick a different colourscheme for each alignment, and use the {\sl
+Colour by ..} submenu to ensure they are all used to colour the complex shown
+in the Jmol window.}
+\exstep{The final step needed to reproduce the shading in Figure
+\ref{mviewalcomplex} is to use the {\sl Colour $\Rightarrow$ By
+Annotation } option in each alignment window to shade the alignment by the
+{\bf Conservation} annotation row. This function was described in section
+\ref{colourbyannotation}.
+
+{\sl Note: Choose a different shading scheme for each
+alignment so that the regions of strong physicochemical conservation are highlighted. This
+kind of shading will reveal conserved regions of interaction between domains
+in the structure.}}
+\exstep{Save your work as a Jalview project and verify that it can be opened again by starting another Jalview Desktop instance, and dragging the saved project into the desktop window.}
+{\sl 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
+bug (see
+\href{http://issues.jalview.org/browse/JAL-1008}{http://issues.jalview.org/browse/JAL-1008}
+for one relating to highlighting of positions in the alignment window).} }
+
+\section{Analysis of alignments}
+\label{alignanalysis}
+Jalview provides support for sequence analysis in two ways. A number of analytical methods are `built-in' and run inside Jalview itself and are mostly accessed from the {\sl Calculate} alignment window menu. Computationally intensive analyses are run outside Jalview via web services - these are typically accessed via the {\sl Web Services} menu, and described in Section \ref{jvwebservices}. In this section, we describe the built-in analysis capabilities common to both the Jalview Desktop and the JalviewLite applet.
\subsection{PCA}
-This calculation creates a spatial representation of the similarities within the current selection or the whole alignment if no selection has been made. After the calculation finishes, a 3D viewer displays the each sequence as a point in 3D 'similarity space'. Sets of similar sequences tend to lie near each other in this space.
-Note: The calculation is computationally expensive, and may fail for very large sets of sequences - because the JVM has run out of memory. See the FAQ on the Jalview web site for instructions on how to increase the memory available to Jalview.
+This calculation creates a spatial representation of the similarities within the current selection or the whole alignment if no selection has been made. After the calculation finishes, a 3D viewer displays the each sequence as a point in 3D `similarity space'. Sets of similar sequences tend to lie near each other in this space.
+Note: The calculation is computationally expensive, and may fail for very large sets of sequences - because the JVM has run out of memory. Memory issues were discussed in Section \ref{memorylimits}.
\subsubsection{What is PCA?}
Principal components analysis is a technique for examining the structure of complex data sets. The components are a set of dimensions formed from the measured values in the data set, and the principle component is the one with the greatest magnitude, or length. The sets of measurements that differ the most should lie at either end of this principle axis, and the other axes correspond to less extreme patterns of variation in the data set.
Labels will be shown for each sequence by toggling the {\sl View $\Rightarrow$ Show Labels} menu option, and the plot background colour changed via the {\sl View $\Rightarrow$ Background Colour..} dialog box. A graphical representation of the PCA plot can be exported as an EPS or PNG image via the {\sl File $\Rightarrow$ Save As $\Rightarrow$ \ldots } submenu.
\exercise{Principle Component Analysis}{
-\exstep{Load the alignment at \textsf{http://www.jalview.org/examples/exampleFile.jar} and select {\sl Edit $\Rightarrow$ Undefine Groups}.
-}
-\exstep{Select the menu option {\sl Calculate $\Rightarrow$ Principle Component Analysis}. A new window will open. Move this window so that the tree, alignment and PCA viewer window are all visible. Try rotating the plot by clicking and dragging the mouse on the plot in the PCA window. Note that clicking on points in the plot will highlight them on the alignment and tree.
-}
+\exstep{Load the alignment at
+\textsf{http://www.jalview.org/examples/exampleFile.jar} and select {\sl Select
+$\Rightarrow$ Undefine Groups}. } \exstep{Select the menu option {\sl Calculate $\Rightarrow$ Principle Component Analysis}. A new window will open. Move this window so that the tree, alignment and PCA viewer window are all visible. Try rotating the plot by clicking and dragging the mouse on the plot in the PCA window. Note that clicking on points in the plot will highlight them on the alignment and tree. }
\exstep{
Click on the tree window. Careful selection of the tree partition location will divide the alignment into a number of groups, each of a different colour. Note how the colour of the sequence ID label matches both the colour of the partitioned tree and the points in the PCA plot.
}
}
\subsection{Trees}
+\label{trees}
Jalview can calculate and display trees, providing interactive tree-based grouping of sequences though a tree viewer. All trees are calculated via the {\sl Calculate $\Rightarrow$ Calculate Tree $\Rightarrow$ \ldots} submenu. Trees can be calculated from distance matrices determined from \% identity or aggregate BLOSUM 62 score using either average distance (UPGMA) or Neighbour joining algorithms. The input data for a tree calculation is either the visible portions of the current selection, or the whole alignment if no selection is present.
-On calculating a tree, a new window opens (Figure \ref{trees1}) which contains the tree. Various display options can be found in the tree window {\sl View} menu, and export options in the {\sl File $\Rightarrow$ Save As} submenu. Newick format is a standard file format for trees which allows them to be exported to other programs. Jalview can also read in external trees in Newick format via the {\sl File $\Rightarrow$ Load Associated Tree} menu option. Leaf names on imported trees will be matched to the associated alignment - unmatched leaves will still be displayed, and can be highlighted using the {\sl View $\Rightarrow$ Show Unlinked Leaves }.
+On calculating a tree, a new window opens (Figure \ref{trees1}) which contains
+the tree. Various display settings can be found in the tree window {\sl View}
+menu, including font, scaling and label display options, and the {\sl File
+$\Rightarrow$ Save As} submenu contains options for image and Newick file
+export. Newick format is a standard file format for trees which allows them to
+be exported to other programs. Jalview can also read in external trees in Newick format via the {\sl File $\Rightarrow$ Load Associated Tree} menu option. Leaf names on imported trees will be matched to the associated alignment - unmatched
+leaves will still be displayed, and can be highlighted using the {\sl View
+$\Rightarrow$ Show Unlinked Leaves} menu option.
\begin{figure}
\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 group them together, select the {\sl Calculate $\Rightarrow$ Sort $\Rightarrow$ By Tree Order} alignment window menu option and the correct tree. The sequences will then be sorted to group them together. The coloured background to the sequence IDs can be removed with {\sl Select $\Rightarrow$ Undefine Groups} from the alignment window menu.
+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 group them together, select the {\sl Calculate $\Rightarrow$ Sort $\Rightarrow$ By Tree Order} alignment window menu option and the correct tree. The sequences will then be sorted according to the leaf order currently shown in the tree view. The coloured background to the sequence IDs can be removed with {\sl Select $\Rightarrow$ Undefine Groups} from the alignment window menu.
\begin{figure}
\begin{center}
%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}
-\parbox[c]{4in}{
-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]{3in}{\centerline{\includegraphics[width=2.75in]{images/pca_vmenu.pdf}
+\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}
-\parbox[c]{5in}{
+\parbox[c]{4in}{
The {\sl View $\Rightarrow$ Associated View $\Rightarrow$ .. } 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 in accord with the colouring and selection state of the newly associated view.
-} \parbox[c]{=2in}{\centerline{ \includegraphics[width=1.3in]{images/pca_fmenu}
-}}
+} \parbox[c]{3in}{\centerline{
+\includegraphics[width=2.5in]{images/pca_vmenu.pdf} }}
\exercise{Trees}{
}
\exstep{Recover the input data for the tree you just calculated. Check the {\sl Edit $\Rightarrow$ Pad Gaps } option is not ticked, and insert one gap in the alignment. Now select {\sl Calculate $\Rightarrow$ Calculate Tree $\Rightarrow$ Neighbour Joining Using BLOSUM62}.
-A warning dialog box {\bf ``Sequence's must be aligned'' } appears because the sequences input to the tree calculation are of different lengths. }
+A warning dialog box {\bf ``Sequences must be aligned'' } appears because the sequences input to the tree calculation are of different lengths. }
-\exstep{Now select {\sl Edit $\RightArrow$ Pad Gaps } and try to perform the tree calculation again - this time a new tree should appear.
+\exstep{Now select {\sl Edit $\Rightarrow$ Pad Gaps } and try to perform the tree calculation again - this time a new tree should appear.
This demonstrates the use of the {\sl Pad Gaps } editing preference, which ensures that all sequences are the same length after editing. }
\subsection{Tree Based Conservation Analysis}
\label{treeconsanaly}
+
+Trees reflect the pattern of global sequence similarity exhibited by the
+alignment or region within the alignment that was used for their calculation.
+The Jalview tree viewer enables sequences to be partitioned into groups based
+on the tree, by clicking within the tree viewer window. Once subdivided, the
+conservation between and within groups can be visually compared in order to
+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 $\Rightarrow$ Automatic
+Annotation $\Rightarrow$ Group Consensus} and {\sl Group Conservation} options)
+can help when working with larger alignments.
+
\exercise{Tree Based Conservation Analysis}{
+\label{consanalyexerc}
\exstep{Load the PF03460 Seed alignment using the sequence fetcher. Colour it with the {\sl Taylor colourscheme}, and apply {\sl Conservation } shading. }
\exstep{Build a Neighbourjoining tree using BLOSUM62 and use the sort submenu to order alignment using the calculated tree.}
\exstep{Select a point on the tree to partition the alignment, and examine the variation in colouring between different groups.
You may find it easier to browse the alignment by if you uncheck the {\sl Show Annotations} view option, and open the overview window to aid navigation.}
\exstep{Try changing the colourscheme to BLOSUM62 (whilst ensuring that {\sl Apply Colour to All Groups} is selected)}
-}
+{\sl Note: You may want to save the alignment and tree as a project file, since
+it is used in the next few exercises. } }
\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 eliminate these sequences from the alignment as an edit operation.
+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, rather than deleted.}.
\begin{figure}
\begin{center}
\includegraphics[width=5.5in]{images/redundancy.pdf}
\end{center}
\label{removeredundancydialog}
-\caption{The Redundancy Removal dialog box opened from the edit menu. Sequences that exceed the current \%age identity threshold and are to be removed are highlighted in black.}
+\caption{The Redundancy Removal dialog box opened from the edit menu. Sequences that exceed the current percentage identity threshold and are to be removed are highlighted in black.}
\end{figure}
\exercise{Remove redundant sequences}{
-\exstep{Re-use or recreate the alignment and tree which you worked with in the tree based conservation analysis exercise (exercise \ref{treeconsanaly})}
-\exstep{Open the Remove Redundancy dialog and adjust the threshold to 90\%. Remove the seuqences that are more than 90\% similar under this alignment.}
+\exstep{Re-use or recreate the alignment and tree which you worked with in the
+tree based conservation analysis exercise (exercise \ref{consanalyexerc})}
+\exstep{Open the Remove Redundancy dialog and adjust the threshold to 90\%. Remove the sequences that are more than 90\% similar under this alignment.}
\exstep{Select the Tree viewer's {\sl View $\Rightarrow$ Show Linked Leaves} option, and note that the removed sequences are now prefixed with a * in the tree view.}
\exstep{Use the [Undo] button on the dialog to recover the sequences. Note that the * symbols disappear from the tree display.}
\exstep{Experiment with the redundancy removal and observe the relationship between the percentage identity threshold and the pattern of unlinked nodes in the tree display.}
}
-\subsection{Other Calculations}
+\subsection{Subdividing the alignment according to specific mutations}
+
+It is often necessary to explore variations in an alignment that may correlate
+with mutations observed in a particular region; for example, sites
+exhibiting single nucleotide polymorphism, or residues involved in substrate
+recognition in an enzyme. One way to do this would be to calculate a tree using
+the specific region, and subdivide it in order to partition the alignment.
+However, calculating a tree can be slow for large alignments, and the tree may be difficult to partition when complex mutation patterns are being analysed. The {\sl Select $\Rightarrow$ Make groups for
+selection } function was introduced to make this kind of analysis easier. When
+selected, it will use the characters in the currently selected region to
+subdivide the alignment. For example, if a single column is selected, then the
+alignment (or each group defined on the alignment) will be divided into groups
+based on the residue or nucleotide found at that position. These new groups are
+annotated with the characters in the selected region, and Jalview's group based
+conservation analysis annotation and colourschemes can then be used to reveal
+any associated pattern of sequence variation across the whole alignment.
+
+\subsection{Automated annotation of Alignments and Groups}
+
+On loading a sequence alignment, Jalview will
+normally\footnote{Automatic annotation can be turned off in the
+{\sl Visual } tab in the {\sl Tools $\Rightarrow$ Preferences } dialog box.}
+calculate a set of automatic annotation rows which are shown below the
+alignment. For nucleotide sequence alignments, only an alignment
+consensus row will be shown, but for amino acid sequences,
+alignment quality (based on BLOSUM62) and physicochemical conservation will
+also be shown. Conservation is calculated according to Livingstone and
+Barton\footnote{{\sl ``Protein Sequence Alignments: A Strategy for the Hierarchical Analysis of Residue Conservation."
+} Livingstone C.D. and Barton G.J. (1993) {\sl CABIOS } {\bf 9}, 745-756}.
+Consensus is the modal residue (or {\tt +} where there is an equal top
+residue). The inclusion of gaps in the consensus calculation can be toggled by
+right-clicking on the the Consensus label and selecting {\sl Ignore Gaps in
+Consensus} from the context menu. Quality is a measure of the inverse
+likelihood of unfavourable mutations in the alignment. Further details on these
+calculations can be found in the on-line documentation.
+
+These annotations can be hidden and deleted but are only created on loading an
+alignment. If they are deleted then the alignment should be saved and reloaded
+to restore them. Jalview provides a toggle to autocalculate a consensus
+sequence upon editing. This is normally left selected but for large alignments
+can be turned off via the {\sl Calculate $\Rightarrow$ Autocalculate
+Consensus} menu option if the interface is too sluggish.
+
+\subsubsection{Group Associated Annotation}
+\label{groupassocannotation}
+Group associated consensus and conservation annotation rows reflect the
+sequence variation within a particular group. Their calculation is enabled
+by selecting the {\sl Group Conservation} or {\sl Group Consensus} options in
+the {\sl View $\Rightarrow$ Automatic Annotation } submenu of the alignment
+window.
+
+\subsubsection{Alignment and Group Sequence Logos}
+\label{seqlogos}
+
+The consensus annotation row that is shown below the alignment can be overlaid
+with a sequence logo that reflects the symbol distribution at each column of
+the alignment. Right click on the Consensus annotation row and select the `Show
+Logo' option to display the Consensus profile for the group or alignment.
+Sequence logos can be enabled by default for all new alignments {\sl via} the
+Visual tab in the Jalview desktop's preferences dialog box.
+
+\exercise{Group conservation analysis}{
+\exstep{Re-use or recreate the alignment and tree which you worked with in the
+tree based conservation analysis exercise (exercise \ref{consanalyexerc})} \exstep{Create a new view, and ensure the annotation panel is displayed, and
+enable the display of {\sl Group Consensus}, and the display of sequence
+logos to make it easier to see the different residue populations within each group.}
+\exstep{Select a column exhibiting about 50\% conservation that lies within the
+central conserved region of the alignment. Subdivide the alignment according to
+this selection using {\sl Select $\Rightarrow$ Make groups for selection}.}
+\exstep{Re-order the alignment according to the new groups that have been
+defined. Click on the group annotation row IDs to select groups exhibiting a
+specific mutation.}
+\exstep{Select another column exhibiting about 50\% conservation
+overall, and subdivide the alignment further. Note that the new groups
+inherit the names of the original groups, allowing you to identify the
+combination of mutations that resulted in the subdivision.
+}
+\exstep{Clear the groups, and try to subdivide the alignment using two
+non-adjacent columns.
+{\sl Hint: You may need to hide the intervening columns
+before you can select both of the columns that you wish to use to subdivide
+the alignment.}}
+\exstep{Switch back to the original view, and experiment with subdividing the tree groups made in the previous excercise.}
+}
-On loading a sequence alignment, Jalview calculates alignment quality, conservation and consensus as alignment annotations. Conservation is calculated according to Livingstone and Barton\footnote{{\sl "Protein Sequence Alignments: A Strategy for the Hierarchical Analysis of Residue Conservation." } Livingstone C.D. and Barton G.J. (1993) {\sl CABIOS } {\bf 9}, 745-756}. Consensus is the modal residue (or {\tt +} where there is an equal top residue). The inclusion of gaps in the consensus calculation can be toggled by right-clicking on the the Consensus label and selecting {\sl Ignore Gaps in Consensus} from the context menu. Quality is a measure of the inverse likelihood of unfavourable mutations in the alignment. Further details on these calculations can be found in the on-line documentation.
+\subsection{Other Calculations}
-These annotations can be hidden and deleted but are only created on loading an alignment. If they are deleted then the alignment should be saved and reloaded to restore them. Jalview provides a toggle to autocalculate a consensus sequence upon editing. This is normally left selected but for large alignments can be turned off via the {\sl Calculate $\Rightarrow$ Autocalculate Consensus} menu option if the interface is too sluggish.
\subsubsection{Pairwise Alignments}
Jalview can calculate optimal pairwise alignments between arbitrary sequences via the {\sl Calculate $\Rightarrow$ Pairwise Alignments\ldots} menu option. Global alignments of all pairwise combinations of the selected sequences are performed and the results returned in a text box.
-\begin{figure}[htbp]
+\begin{figure}[]
\begin{center}
\includegraphics[width=4in]{images/pairwise.pdf}
\caption{{\bf Pairwise alignment of sequences.} Pairwise alignments of three selected sequences are shown in a textbox.}
\end{center}
\end{figure}
+\pagebreak[2]
+
+\section{Webservices}
+\label{jvwebservices}
+The term ``Webservices'' refers to a variety of data exchange
+mechanisms based on HTTP.\footnote{HTTP: Hyper-Text Transfer Protocol.}
+
+\parbox[c]{4.5in}{Jalview can exploit public webservices to access databases
+remotely, and also submit data to public services by opening pages with your web browser. These types of
+services are `one-way', {\sl i.e.} data is either sent to the webservice or
+retrieved from it by Jalview. The desktop application can also interact
+with `two-way' remote analysis services in order to offload computationally
+intensive tasks to High Performance Computing facilities. Most of these two-way
+services are provided by {\bf Ja}va {\bf B}ioinformatics {\bf A}nalysis {\bf
+W}eb {\bf S}ervice (JABAWS) servers\footnote{See
+http://www.compbio.dundee.ac.uk/jabaws for more information and to download
+your own server.}, which provides an easily installable system for performing
+a range of bioinformatics analysis tasks. }
+\parbox[c]{1.75in}{\includegraphics[width=1.65in]{images/wsmenu.pdf}}
+
+\subsection{One way web services}
+
+There are three types of one way service in jalview. Database services,
+which were introduced in in Section \ref{fetchseq}, provide sequence and
+alignment data. They can also be used to add sequence IDs to an alignment
+imported from a local file, prior to further annotation retrieval, as described
+in Section \ref{featuresfromdb}. A second type of one way service is provided
+by Jalview's DAS sequence feature retrieval system, which is described
+in Section \ref{dasfretrieval}. The final type of one way service are sequence
+and ID submission services, exemplified by the `Envision2 Services' provided
+by the ENFIN Consortium\footnote{ENFIN is the European Network for Functional
+INtegration. Please see http://www.enfin.org for more information. }.
+
+\subsubsection{One-way submission services}
+Jalview can use the system's web browser to submit sets of sequences and
+sequence IDs to web based applications. Single sequence IDs can be passed to
+a web site using the user definable URL links listed under the {\sl
+Links} submenu of the sequence ID popup menu. These are configured
+in the {\sl Connections} tab of the {\sl Preferences} dialog box.
+
+The Envision2 services presented in the webservice menu provides are the first
+example of one way services where multiple sequences or sequence IDs can be
+sent. The {\sl Web services $\Rightarrow$ Envision2 Services} menu entry
+provides two sub-menus that enable you to submit the sequences or IDs
+associated with the alignment or just the currently selected sequences to one
+of the Envision2 workflows. Selecting any one will open a new browser window on
+the Envision2 web application. The menu entries and their tooltips provide
+details of the Envision2 workflow and the dataset set that will be submitted
+({\sl i.e.} the database reference type, or associated sequence subset). Please
+note, due to technical limitations, Jalview can currently only submit small
+numbers of sequences to the workflows - if no sequence or ID submissions are
+presented in the submenus, then try to select a smaller number of sequences to
+submit.
+
+\subsection{Remote Analysis Web Services}
+Remote analysis services enable Jalview to use external computational
+facilities. There are curently three types of service - multiple sequence
+alignment, protein secondary structure prediction, and alignment analysis.
+Many of these are provided by JABA servers, which are described at the end of
+this section. In all cases, Jalview will construct a job based on the alignment
+or currently selected sequences, ask the remote server to run the job, monitor
+status of the job and, finally, retrieve the results of the job and display
+them. The Jalview user is kept informed of the progress of the job through a
+status window.
+
+Currently, web service jobs and their status windows are not stored in Jalview
+Project Files\footnote{This may be rectified in future versions.}, so it is
+important that you do not close Jalview whilst a job is running. It is also
+essential that you have a continuous network connection in order to
+successfully use Web Services from Jalview, since it periodically checks the
+progress of running jobs.
+
+
+\subsection{JABA Web Services for sequence alignment and analysis}
+JABA stands for ÔJAva Bioinformatics AnalysisÕ, which is a system developed by
+Peter Troshin and Geoff Barton at the University of Dundee for running
+computationally intensive bioinformatics analysis programs. A JABA installation
+typically provides a range of JABA web services (JABAWS) for use by other
+programs, like Jalview.
+
+Exercises in the remainder of this section will demonstrate the simplest way of
+installing JABA on your computer, and configuring Jalview so it can access the JABA services. If you
+need any further help or more information about the services, please go to the
+\href{http://www.compbio.dundee.ac.uk/jabaws}{JABAWS home page}.
+%% \subsubsection{Aims}
+%% \begin{list}{$\bullet$}{}
+%% \item Gain experience using the different alignment services provided by
+% JABA
+%%\item Learn about the way that Jalview stores user presets for JABA services
+%%\item Learn how to install JABA services and configure Jalview to access them
+%%\end{list}
+
+\subsection{Changing the Web Services menu layout}
+\label{changewsmenulayout}
+If you are working with a lot of different JABA services, you may wish to
+change the way Jalview lays out the web services menu. You can do this from the Web Services tab of the Preferences dialog box.
+
+\exercise{Changing the Layout of the Web Services Menu}{
+\label{changewsmenulayoutex}
+\exstep{Make sure you have loaded an alignment into Jalview, and examine the
+current layout of the alignment windowÕs Web Services menu.}
+\exstep{Open the preferences dialog box and select the web services tab.}
+\exstep{Ensure the ÔEnable JABAWS servicesÕ checkbox is selected, and unselect
+the ÔEnable Enfin ServicesÕ checkboxes.}
+\exstep{Hit ÔRefresh servicesÕ to update the web service menu -- once the
+progress bar has completed, open the web services menu to view the changes.}
+\exstep{Unselect the ÔIndex by hostÕ checkbox and refresh the services once again.
+
+{\sl Observe the way the layout of the JABAWS Alignment submenu changes.}
+}
+\exstep{Do the same with the ÔIndex by typeÕ checkbox.}
+}
+
+Jalview provides these options for configuring the layout of the web services
+menu because different Jalview users may have access to a different number of
+JABA services, and each will have their own preference regarding the layout of
+the menu.
+
+\begin{figure}[htbc]
+\begin{center}
+\includegraphics[width=3in]{images/jvjabawsconfig.pdf}
+\caption{{\bf The Jalview Web Services preferences panel.} Options are provided
+for configuring the list of JABA servers that Jalview will use, enabling and
+disabling Enfin services, and configuring the layout of the web services
+menu.}
+\end{center}
+\end{figure}
-\section{Web Services}
-Web services are a technology which allows the Jalview application to access databases remotely and offload computationally intensive tasks to High Performacne Computing facilities {\sl via} the internet. Access to remote database resources are described in other sections. They include sequence retrieval (in section \ref{fetchseq}), database reference retrieval (in section \ref{fetchdbrefs}) and sequence feature retrieval with DAS, which is described in \ref{das}. The next few sections describe the computational analysis services available to Jalview.
-
-\subsection{Remote Analysis Services}
-Jalview will construct a job, ask the remote server to run the job, monitor status of the job and, finally, retrieve the results of the job and display them. The Jalview user is kept informed of the progress of the job through a status window.
-
-Currently, web service jobs and their status windows are not stored in Jalview Project Files\footnote{This may be rectified in future versions.}, so it is important that you do not close Jalview whilst a job is running. It is also essential that you have a continuous network connection in order to successfully use Web Services from Jalview, since it periodically checks the progress of running jobs.
-
-\subsection{Multiple Sequence Alignment}
-
-Sequences can be aligned using any of three algorithms: ClustalW\footnote{{\sl "CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position specific gap penalties and weight matrix choice."} Thompson JD, Higgins DG, Gibson TJ (1994) {\sl Nucleic Acids Research} {\bf 22}, 4673-80}, Muscle\footnote{{\sl "MUSCLE: a multiple sequence alignment method with reduced time and space complexity"} Edgar, R.C. (2004) {\sl BMC Bioinformatics} {\bf 5}, 113} or MAFFT\footnote{{\sl "MAFFT: a novel method for rapid multiple sequence alignment based on fast Fourier transform"} Katoh, K., Misawa, K., Kuma, K. and Miyata, T. (2002) {\sl Nucleic Acids Research} {\bf 30}, 3059-3066. and {\sl "MAFFT version 5: improvement in accuracy of multiple sequence alignment"} Katoh, K., Kuma, K., Toh, H. and Miyata, T. (2005) {\sl Nucleic Acids Research} {\bf 33}, 511-518.} Of these, ClustalW is the slowest but is historically widely used. Muscle is fast and probably the most accurate for smaller alignments and MAFFT is probably the best for large alignments.
-
-
-To run an alignment web service, select the appropriate method from the {\sl Web Service $\Rightarrow$ Alignment $\Rightarrow$ \ldots} submenu (Figure \ref{webservices}). A progress window will appear giving information about the job and any errors that occur. After successful completion of the job, a new window is opened with the results, in this case an alignment.
+\subsection{Running your own JABA server}
+You can download and run JABA on your own machine using the ÔVMWareÕ or
+VirtualBox virtual machine environments. If you would like to learn how to do
+this, there are full instructions at the
+\href{http://www.compbio.dundee.ac.uk/jabaws/}{JABA web site}.
+
+\exercise{Installing a JABA Virtual Machine on your computer}{
+\label{jabawsvmex}{\sl This tutorial will demonstrate the simplest way of
+installing JABA on your computer, and configuring Jalview so it can access the JABA services.
+
+{\bf Prerequisites}
+
+You will need a copy of VMWare Player/Workstation/Fusion on your machine.
+}
+
+\exstep{If you do not have VMWare player installed, download it from
+www.vmware.com (this takes a few minutes Ð you will need to register and wait
+for an email with a download link).}
+\exstep{Download the JABA virtual appliance archive called Ôjaba-vm.zipÕ from
+\textsf{http://www.compbio.dundee.ac.uk/jabaws/archive/jabaws-vm.zip}
+
+WARNING: This is large ($~$300MB) and will take some time to download.
+}
+\exstep{Unpack the archiveÕs contents to a place on your machine with at least
+2GB of free space.
+
+(On windows, right click on the archive, and use the ÔExtract archive..Õ option).
+}
+\exstep{Open the newly extracted directory and double click the VMWare virtual
+machine configuration file (jabaws.vcf). This will launch the VMWare player.
+}
+\exstep{Once VMWare player has started up, it may ask the question Ôdid you move or copy
+this virtual appliance?Õ Ð select ÔcopyÕ.}
+\exstep{You may be prompted to download the VMWare linux tools Ð these are not
+necessary, so close the window or click on ÔlaterÕ}
+\exstep{You may also be prompted to install support for one or more devices (USB
+or otherwise) Ð say no to these options.}
+\exstep{Once the machine has loaded, it will display a series of IP addresses
+for the different services provided by the VM. Make a note of the JABAWS URL Ð
+this will begin with Ôhttp:Õ and end with Ô/jabawsÕ.}
+}
+
+\exercise{Configuring Jalview to access your new JABAWS virtual appliance}{
+\label{confnewjabawsappl}
+\exstep{Start Jalview (If you have not done so already).}
+\exstep{Enable the Jalview Java Console by selecting its option from the Tools
+menu.
+
+{\sl Alternately, use the System Java console if you have
+configured it to open when Jalview is launched, via your systemÕs Java
+preferences (under the ÔAdvancedÕ tab on Windows).}}
+\exstep{Open the preferences dialog and locate the Web Services preferences.}
+\exstep{Add the URL for the new JABAWS server you started in Exercise
+\ref{jabawsvmex} to the list of JABAWS urls using the ÔNew Service
+URLÕ button.}
+\exstep{You will be asked if you want to test the service. Hit ÔYesÕ to do this
+Ð- you should then see some output in the console window.
+
+{\sl Take a close look at the output in the console. What do you think is
+happening?}
+}
+\exstep{Hit {\sl OK} to save your preferences Ð- you have now added a new JABA
+service to Jalview!}
+\exstep{Try out your new JABA services by loading the ferredoxin sequences from
+http://www.jalview.org/tutorial/alignment.fa}
+\exstep{Launch an alignment using one
+of the JABA methods provided by your server. It will be listed under the JABAWS Alignment submenu of the web services menu on the alignment window.
+
+{\sl Note: You can watch the JABA VM appliance's process working by opening the
+process monitor on your system. (On Windows XP, this involves right-clicking the
+system clock and opening the task manager Ð- then selecting the ÔProcessesÕ tab
+and sort by CPU).}
+}
+}
+
+
+\subsubsection{Resetting the JABA services setting to their defaults}
+Once you have configured a JABAWS server and selected the `OK' button of the
+preferences menu, the settings will be stored in your Jalview preferences file,
+along with any preferences regarding the layout of the web services menu. If
+you should ever need to reset the JABAWS server list to its defaults, use the
+`Reset Services' button on the Web Services preferences panel.
+
+\section{Multiple Sequence Alignment}
+\label{msaservices}
+Sequences can be aligned using a range of algorithms provided by JABA web
+services. These include ClustalW,\footnote{{\sl ``CLUSTAL W: improving the
+sensitivity of progressive multiple sequence alignment through sequence weighting, position specific gap penalties and weight matrix choice."} Thompson JD, Higgins DG, Gibson TJ (1994) {\sl Nucleic
+Acids Research} {\bf 22}, 4673-80} Muscle,\footnote{{\sl ``MUSCLE: a multiple
+sequence alignment method with reduced time and space complexity"} Edgar, R.C.
+(2004) {\sl BMC Bioinformatics} {\bf 5}, 113} MAFFT,\footnote{{\sl ``MAFFT: a
+novel method for rapid multiple sequence alignment based on fast Fourier
+transform"} Katoh, K., Misawa, K., Kuma, K. and Miyata, T. (2002) {\sl
+Nucleic Acids Research} {\bf 30}, 3059-3066. and {\sl ``MAFFT version 5:
+improvement in accuracy of multiple sequence alignment"} Katoh, K., Kuma, K.,
+Toh, H. and Miyata, T. (2005) {\sl Nucleic Acids Research} {\bf 33}, 511-518.}
+ProbCons\footnote{PROBCONS: Probabilistic Consistency-based Multiple Sequence Alignment.
+Do, C.B., Mahabhashyam, M.S.P., Brudno, M., and Batzoglou, S.
+(2005) {\sl Genome Research} {\bf 15} 330-340.} and T-COFFEE.\footnote{T-Coffee:
+A novel method for multiple sequence alignments. (2000) Notredame,Higgins and Heringa {\sl JMB} {\bf 302} 205-217} Of these,
+T-COFFEE is the slowest, but also the most accurate. ClustalW is historically the most widely used. Muscle is faster than ClustalW and probably the most accurate for smaller alignments and MAFFT is probably the best for large
+alignments\footnote{Clustal Omega, which was released in 2011, is arguably the
+fastest and most accurate tool for protein multiple alignment, and is
+included in JABAWS 2.0.}.
+
+
+To run an alignment web service, select the appropriate method from the {\sl
+Web Service $\Rightarrow$ Alignment $\Rightarrow$ \ldots} submenu (Figure
+\ref{webservices}). For each service you may either perform an alignment with
+default settings, use one of the available presets, or customise the parameters
+with the `{\sl Edit and Run ..}' dialog box. Once the job is submitted, a
+progress window will appear giving information about the job and any errors that
+occur. After successful completion of the job, a new window is opened with the
+results, in this case an alignment. By default, the new alignment will be
+ordered in the same way as the input sequences; however, many alignment programs
+re-order the input to place homologous sequences close together. This ordering
+can be recovered using the `Original ordering' entry within the {\sl Calculation
+$\Rightarrow$ Sort } sub menu.
\begin{figure}[htbp]
\begin{center}
\parbox[c]{1.5in}{\includegraphics[width=1.5in]{images/ws1.pdf}}
\parbox[c]{2.5in}{\includegraphics[width=2.5in]{images/ws2.pdf}}
\parbox[c]{2in}{\includegraphics[width=2in]{images/ws3.pdf}}
-\caption{{\bf Multiple alignment via web services} The appropriate method is selected from the menu (left), a status box appears (center), and the results appear in a new window (right)}
+\caption{{\bf Multiple alignment via web services} The appropriate method is
+selected from the menu (left), a status box appears (centre), and the results
+appear in a new window (right)}
\label{webservices}
\end{center}
\end{figure}
\subsubsection{Realignment}
-The re-alignment option (only supported by ClustalW currently) submits the current selection to the alignment service complete with existing gaps. Use this option to align additional sequences to an existing alignment (a simple form of profile alignment).
+The re-alignment option is currently only supported by ClustalW. When performing a re-alignment, Jalview submits the current selection to the alignment service complete with any existing gaps. This approach is useful when one wishes to align additional sequences to an existing alignment without any further optimisation to the existing alignment. The Re-alignment service provided by ClustalW in this case is effectively a simple form of profile alignment.
-\subsubsection{Alignments of sequences including hidden regions}
+\subsubsection{Alignments of sequences that include hidden regions}
-If the view or selected region that is submitted for alignment contains hidden regions, then only the visible sequences will be submitted to the service. Furthermore, each contiguous segment of sequences will be aligned independently (resulting a number of alignment 'subjobs' appearing in the status window). Finally, the results of each subjob will be concatenated with the hidden regions in the input data prior to their display in a new window. This approach ensures that 1) hidden column boundaries in the input data are preserved in the resulting alignment - in a similar fashion to the constraint that hidden columns place on alignment editing (see section \ref{lockededits}). 2) hidden columns can be used to preserve existing parts of an alignment whilst the visible parts are locally refined.
+If the view or selected region that is submitted for alignment contains hidden regions, then only the visible sequences will be submitted to the service. Furthermore, each contiguous segment of sequences will be aligned independently (resulting a number of alignment `subjobs' appearing in the status window). Finally, the results of each subjob will be concatenated with the hidden regions in the input data prior to their display in a new window. This approach ensures that 1) hidden column boundaries in the input data are preserved in the resulting alignment - in a similar fashion to the constraint that hidden columns place on alignment editing (see Section \ref{lockededits}). 2) hidden columns can be used to preserve existing parts of an alignment whilst the visible parts are locally refined.
\exercise{Multiple Sequence Alignment}{
-\exstep{ Close all windows and open the alignment at {\sf http://www.jalview.org/tutorial/unaligned.fa}. Select {\sl Web Service $\Rightarrow$ Alignment $\Rightarrow$ Muscle Multiple Protein Sequence Alignment}. A window will open giving the job status. After a short time, a second window will open with the results of the alignment.}
-\exstep{Select the first sequence set by clicking on the window and try running ClustalW and MAFFT (from the {\sl Web Services $\Rightarrow$ Alignment} menu) on the same initial alignment. Compare them and you should notice small differences.
-}
+\exstep{ Close all windows and open the alignment at {\sf
+http://www.jalview.org/tutorial/unaligned.fa}. Select {\sl
+Web Service $\Rightarrow$ Alignment $\Rightarrow$ Default Settings} for Muscle. A window will open giving the job status. After a short time, a second window will open with the results of the alignment.} \exstep{Select the first sequence set by clicking on the window and try running ClustalW and MAFFT (from the {\sl Web Services $\Rightarrow$ Alignment} menu) on the same initial alignment. Compare them and you should notice small differences. }
\exstep{Select the last three sequences in the MAFFT alignment, and de-align them with {\sl Edit $\Rightarrow$ Remove All Gaps}. Press [ESC] to deselect them and then submit the view for re-alignment with ClustalW.}
\exstep{Use [CTRL]-Z to recover the alignment of the last three sequences in the MAFFT alignment. Once the ClustalW re-alignment has completed, compare the results of re-alignment of the three sequences with their alignment in the original MAFFT result.}
-\exstep{Select columns 60 to 125 in the original MAFFT alignment and hide them. Select {\sl Web Services $\RightArrow$ Alignment $\RightArrow$ MAFFT} to submit the visible portion of the alignment to MAFFT. When the web service job pane appears, note that there are now two alignment job status panes shown in the window.}
+\exstep{Select columns 60 to 125 in the original MAFFT alignment and hide them. Select {\sl Web Services $\Rightarrow$ Alignment $\Rightarrow$ MAFFT} to submit the visible portion of the alignment to MAFFT. When the web service job pane appears, note that there are now two alignment job status panes shown in the window.}
\exstep{When the MAFFT job has finished, compare the alignment of the N-terminal visible region in the result with the corresponding region of the original alignment. If you wish, select and hide a few more columns in the N-terminal region, and submit the alignment to the service again and explore the effect of local alignment on the non-homologous parts of the N-terminal region.}
}
-\subsection{Secondary Structure Prediction}
+\subsection{Customising the parameters used for alignment}
-Secondary structure prediction is performed using the Jpred\footnote{{\sl "Jpred: A Consensus Secondary Structure Prediction Server"} Cuff, J. A., Clamp, M. E., Siddiqui, A. S., Finlay, M. and Barton, G. J. (1998) {\sl Bioinformatics} {\bf 14}, 892-893} server at the University of Dundee\footnote{http://www.compbio.dundee.ac.uk/~www-jpred/}. The behaviour of this calculation depends on the current selection:
+JABA web services allow you to vary the parameters used when performing a
+bioinformatics analysis. For JABA alignment services, this means you are
+usually able to modify the following types of parameters:
+\begin{list}{$\bullet$}{}
+\item Amino acid or nucleotide substitution score matrix
+\item Gap opening and widening penalties
+\item Types of distance metric used to construct guide trees
+\item Number of rounds of re-alignment or alignment optimisation
+\end{list}
+
+
+\subsubsection{Getting help on the parameters for a service}
+Each parameter available for a method usually has a short description, which
+jalview will display as a tooltip, or as a text pane that can be opened under
+the parameterÕs controls. In the parameter shown in Figure
+\ref{clustalwparamdetail}, the description was opened by selecting the button on the left hand side. Online help for the
+service can also be accessed, by right clicking the button and selecting a URL
+from the pop-up menu that will open.
+
+\begin{figure}[htbp]
+\begin{center}
+\includegraphics[width=2.5in]{images/clustalwparamdetail.pdf}
+\caption{{\bf ClustalW parameter slider detail}. From the ClustalW {\sl Edit &
+Run ..} dialog box. }
+\label{clustalwparamdetail}
+\end{center}
+\end{figure}
+
+\subsection{Alignment Presets}
+The different multiple alignment algorithms available from JABA vary greatly in
+the number of adjustable parameters, and it is often difficult to identify what
+are the best values for the sequences that you are trying to align. For these
+reasons, each JABA service may provide one or more presets Ð which are
+pre-defined sets of parameters suited for particular types of alignment
+problem. For instance, the Muscle service provides the following presets:
+\begin{list}{$\bullet$}{}
+\item Huge
+\item Protein alignments (fastest speed)
+\item Nucleotide alignments (fastest speed)
+\end{list}
+
+The presets are displayed in the JABA web services submenu, and can also
+accessed from the parameter editing dialog box, which is opened by selecting
+the Ô{\sl Edit and run..}Õ option from the web serviceÕs menu. If you have used
+a preset, then it will be mentioned at the beginning of the job status file shown
+in the web service job progress window.
+
+\subsubsection{Alignment Service Limits}
+Multiple alignment is a computationally intensive calculation. Some JABA server
+services and service presets only allow a certain number of sequences to be
+aligned Ð the precise number will depend on the server that you are using to
+perform the alignment. Should you try to submit more sequences than a service
+can handle, then an error message will be shown informing you of the maximum
+number allowed by the server.
+
+\subsection{User defined Presets}
+Jalview allows you to create your own presets for a particular service. To do
+this, select the `{\sl Edit parameters and run ..}' option for your service,
+which will open a parameter editing dialog box like the one shown in Figure
+\ref{jwsparamsdialog}.
+
+The top row of this dialog allows you to browse the existing presets, and
+when editing a parameter set, allows you to change its nickname. As you
+adjust settings, buttons will appear at the top of the parameters dialog that
+allow you to create, update, delete or undo changes to the currently selected
+user preset. In addition to the parameter set name, you can also provide a short
+description for the parameter set, which will be shown in the tooltip for the
+parameter set's entry in the web services menu.
+
+\begin{figure}[htbc]
+\center{
+\includegraphics[width=3in]{images/jvaliwsparamsbox.pdf}
+\caption{{\bf Jalview's JABA alignment service parameter editing dialog box}.}
+\label{jwsparamsdialog} }
+\end{figure}
+
+\subsubsection{Saving parameter sets}
+When creating a custom parameter set, you will be asked for a file name to save
+it. The location of the file is recorded in the Jalview user preferences in the
+same way as a custom alignment colourscheme, so when Jalview is launched again,
+it will show your custom preset amongst the options available for running the
+JABA service.
+
+%
+% \exercise{Creating and using user defined presets}{\label{createandusepreseex}
+% \exstep{Import the file at
+% \textsf{http://www.jalview.org/tutorial/fdx\_unaligned.fa} into jalview.}
+% \exstep{Use the `{\slDiscover Database Ids}' function to recover the PDB cross
+% references for the sequences.}
+% \exstep{Align the sequences using the default ClustalW parameters.}
+% \exstep{Use the `{\sl Edit and run..}'
+% option to open the ClustalW parameters dialog box, and create a new preset using
+% the following settings:
+% \begin{list}{$\bullet$}{}
+% \item BLOSUM matrix (unchanged)
+% \item Gap Opening and End Gap penalties = 0.05
+% \item Gap Extension and Separation penalties = 0.05
+% \end{list}
+%
+% As you edit the parameters, buttons will appear on the dialog box
+% allowing you revert your changes or save your settings as a new parameter
+% set.
+%
+% Before you save your settings, remeber to give them a meaningful name by editing
+% the text box at the top of the dialog box.
+% }
+% \exstep{Repeat the alignment using your new parameter set by selecting it from
+% the {\sl ClustalW Presets menu}.}
+% \exstep{These sequences have PDB structures associated with them, so it is
+% possible to compare the quality of the alignments.
+%
+% Use the {\sl View all {\bf N}
+% structures} option to calculate the superposition of 1fdn on 1fxd for both
+% alignments (refer to section \ref{superposestructs} for instructions). Which
+% alignment gives the best RMSD ? }
+% \exstep{Apply the same alignment parameter settings to the example alignment
+% (available from \textsf{http://www.jalview.org/examples/uniref50.fa}).
+%
+% Are there differences ? If not, why not ?
+% }
+% }
+
+
+\section{Protein Secondary Structure Prediction}
+\label{protsspredservices}
+Protein secondary structure prediction is performed using the
+Jpred\footnote{{\sl ``The Jpred 3 Secondary Structure Prediction Server''} Cole, C., Barber, J. D. and Barton, G. J. (2008) {\sl Nucleic Acids Research} {\bf 36}, (Web Server Issue) W197-W201
+
+{\sl ``Jpred: A Consensus Secondary Structure Prediction Server''} Cuff, J. A.,
+Clamp, M. E., Siddiqui, A. S., Finlay, M. and Barton, G. J. (1998) {\sl
+Bioinformatics} {\bf 14}, 892-893} server at the University of
+Dundee\footnote{http://www.compbio.dundee.ac.uk/www-jpred/}. The behaviour of
+this calculation depends on the current selection:
\begin{list}{$\circ$}{}
-\item If nothing is selected, and the displayed sequences appear to be aligned, then a JNet prediction will be run for the first sequence in the alignment, using the current alignment. Otherwise the first sequence will be submitted for prediction.
+\item If nothing is selected, Jalview will check the length of each alignment row to determine if the visible sequences in the view are aligned.
+\begin{list}{-}{}
+ \item If all rows are the same length (often due to the application of the {\sl Edit $\Rightarrow$ Pad Gaps} option), then a JNet prediction will be run for the first sequence in the alignment, using the current alignment as the profile to use for prediction.
+ \item Otherwise, just the first sequence will be submitted for a full JNet prediction.
+\end{list}
\item If just one sequence (or a region on one sequence) has been selected, it will be submitted to the automatic JNet prediction server for homolog detection and prediction.
-\item If a set of sequences are selected, and they appear to be aligned, then the alignment will be used for a Jnet prediction on the first sequence in the set (that is, the one that appears first in the alignment window).
+\item If a set of sequences are selected, and they appear to be aligned using the same criteria as above, then the alignment will be used for a Jnet prediction on the first sequence in the set (that is, the one that appears first in the alignment window).
\end{list}
-
-Jpred is launched in the same way as the other web services. Select {\sl Web Services $\Rightarrow$ Secondary Structure Prediction $\Rightarrow$ JNet Secondary Structure Prediction} 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.
+Jpred is launched in the same way as the other web services. Select {\sl Web
+Services $\Rightarrow$ Secondary Structure Prediction $\Rightarrow$ JNet
+Secondary Structure Prediction} 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.
\begin{figure}[htbp]
\begin{center}
\subsubsection{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 may\footnote{This, of course, cannot be guaranteed, but the profile calculated by JNet will at least be different.} result in a more reliable secondary structure prediction either side of the insertion. 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.
+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 may result in a more reliable\footnote{This, of course, cannot be guaranteed, but the profile
+calculated by JNet will at least be different.} secondary structure prediction
+either side of the insertion. 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}
}
\exstep{
Select the second sequence prediction, and copy and paste it into the first prediction window. You can now compare the two predictions. Jnet secondary structure prediction annotation are examples of {\bf sequence associated alignment annotation}.
-% which is described in section \ref{seqassocannot} below.
+% which is described in Section \ref{seqassocannot} below.
}
\exstep{
Select and hide some columns in one of the profiles that were returned from the JNet service, and then submit the profile for prediction again.
}
\exstep{
-When you get the result, verify that the prediction has not been made for the hidden parts of the profile, and that the JPred reliability scores differ from the prediction made on the full profile.
+When you get the result, verify that the prediction has not been made for the
+hidden parts of the profile, and that the JPred reliability scores differ from the prediction made on the full profile.
{\sl Note: you may want to keep this data for use in exercise \ref{viewannotfileex}.}
}
\label{featannot}
Features and annotations are additional information that is overlaid on the sequences and the alignment. Generally speaking, annotations are associated with columns in the alignment. Features are associated with specific residues in the sequence.
-Annotations are rendered below the alignment, in the annotation panel, and often reflect properties of the alignment as a whole. The conservation, consensus and quality scores are examples of dynamic annotation. As the alignment changes, these annotations will change along with it. Conversely, features are properties of the individual sequences. They do not change with the alignment, but are shown mapped on to specific residues within the alignment.
+Annotations are rendered below the alignment, in the annotation panel, and often reflect properties of the alignment as a whole. The conservation, consensus and quality scores are examples of dynamic annotation. As the alignment changes, these annotations will change along with it. Conversely, sequence features are properties of the individual sequences. They do not change with the alignment, but are shown mapped on to specific residues within the alignment.
Features and annotation can be interactively created, or retrieved from external data sources. DAS (the Distributed Annotation System) is the primary source of sequence features, whilst webservices like JPred (see \ref{jpred} above) can be used to analyse a given sequence or alignment and generate annotation for it.
\subsection{Creating sequence features}
-Sequence features can be created by simply selecting the area in a sequence (or sequences) to form the feature and selecting {\sl Selection $\Rightarrow$ Create Sequence Feature } from the right-click context menu (Figure \ref{features}). A dialogue box allows the user to customise the feature with respect to name, group, and colour. The feature is then associated with the sequence. Moving the mouse over a residue associated with a feature brings up a tool tip listing all features associated with the residue.
+Sequence features can be created simply by selecting the area in a sequence (or sequences) to form the feature and selecting {\sl Selection $\Rightarrow$ Create Sequence Feature } from the right-click context menu (Figure \ref{features}). A dialogue box allows the user to customise the feature with respect to name, group, and colour. The feature is then associated with the sequence. Moving the mouse over a residue associated with a feature brings up a tool tip listing all features associated with the residue.
\begin{figure}[htbp]
\begin{center}
\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 can be necessary to customise the display. Jalview allows the display, colour, rendering order and transparency of features to be modified via the {\sl View $\Rightarrow$ Feature Settings\ldots} menu option. This brings up a dialogue window (Figure \ref{custfeat}) which allows the visibility of individual feature types to be selected, colours changed (by clicking on the colour of each sequence feature type) and the rendering order modified by dragging feature types to a new position in the list. Dragging the slider alters the transparency of the feature rendering.
+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
+via the {\sl View $\Rightarrow$ Feature Settings\ldots} menu option. This
+brings up a dialogue window (Figure \ref{custfeat}) which allows the
+visibility of individual feature types to be selected, colours changed (by
+clicking on the colour of each sequence feature type) and the rendering order
+modified by dragging feature types to a new position in the list. Dragging the
+slider alters the transparency of the feature rendering. The Feature
+Settings dialog also includes functions for more advanced feature shading
+schemes and buttons for sorting the alignment according to the distribution of
+features. These capabilities are described further in sections
+\ref{featureschemes} and \ref{featureordering}.
\begin{figure}[htbp]
\begin{center}
\includegraphics[width=4in]{images/features4.pdf}
-\caption{{\bf Multiple sequence features.} An alignment with many sequence features overlaid }
+\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}
\end{center}
\end{figure}
+\subsection{Sequence Feature File Formats}
+
+Jalview supports the widely used GFF tab delimited format\footnote{see
+http://www.sanger.ac.uk/resources/software/gff/spec.html} and its own Jalview
+Features file format for the import of sequence annotation. Features and
+alignment annotation are also extracted from other formats such as Stockholm,
+and AMSA. URL links may also be attached to features. See the online
+documentation for more details of the additional capabilities of the jalview
+features file.
+
\exercise{Creating features}{
\exstep{Open the alignment at \textsf{http://www.jalview.org/tutorial/alignment.fa}. We know that the Cysteine residues at columns 97, 102, 105 and 135 are involved in iron binding so we will create them as features. Navigate to column 97, sequence 1. Select the entire column by clicking in the ruler bar. Then right-click on the selection to bring up the context menu and select {\sl Selection $\Rightarrow$ Create Sequence Feature}. A dialogue box will appear.
}
\exstep{
-Enter a suitable Sequence Feature Name (e.g. "Iron binding site") in the appropriate box. Click on the Feature Colour bar to change the colour if desired, add a short description ("One of four Iron binding Cysteines") and press OK. The features will then appear on the sequences.
-}
-\exstep{Roll the mouse cursor over the new features. Note that the position given in the tool tip is the residue number, not the column number. To demonstrate that there is one feature per sequence, clear all selections by pressing [ESC] then insert a gap in sequence 3 at position 95. Roll the mouse over the features and you will see that the feature has moved with the sequence. Delete the gap you created.
+Enter a suitable Sequence Feature Name (e.g. ``Iron binding site") in the
+appropriate box. Click on the Feature Colour bar to change the colour if
+desired, add a short description (``One of four Iron binding Cysteines") and press OK. The features will then appear on the sequences. } \exstep{Roll the mouse cursor over the new features. Note that the position given in the tool tip is the residue number, not the column number. To demonstrate that there is one feature per sequence, clear all selections by pressing [ESC] then insert a gap in sequence 3 at position 95. Roll the mouse over the features and you will see that the feature has moved with the sequence. Delete the gap you created.
}
\exstep{
Add a similar feature to column 102. When the feature dialogue box appears, clicking the Sequence Feature Name box brings up a list of previously described features. Using the same Sequence Feature Name allows the features to be grouped.}
-\exstep{Select {\sl View $\Rightarrow$ Feature Settings\ldots} from the alignment window menu. The Sequence Feature Settings window will appear. Move this so that you can see the features you have just created. Click the check box for "Iron binding site" under {\sl Display} and note that display of this feature type is now turned off. Click it again and note that the features are now displayed. Close the sequence feature settings box by clicking {\sl OK} or {\sl Cancel}.}
-}
+\exstep{Select {\sl View $\Rightarrow$ Feature Settings\ldots} from the
+alignment window menu. The Sequence Feature Settings window will appear. Move
+this so that you can see the features you have just created. Click the check
+box for ``Iron binding site" under {\sl Display} and note that display of this
+feature type is now turned off. Click it again and note that the features are
+now displayed. Close the sequence feature settings box by clicking {\sl OK} or
+{\sl Cancel}.} }
\subsection{Creating user defined annotation}
-Annotations are properties that apply to the alignment as a whole and are visualised on rows in the annotation panel.
+Annotations are properties that apply to the alignment as a whole and are visualized on rows in the annotation panel.
To create a new annotation row, right click on the annotation label panel and select the {\sl Add New Row} menu option (Figure \ref{newannotrow}). A dialogue box appears. Enter the label to use for this row and a new row will appear.
\begin{figure}[htbp]
\end{figure}
\exercise{Annotating alignments}{
-\exstep{Load the alignment at \textsf{http://www.jalview.org/tutorial/alignment.fa}. Right-click on the annotation label for {\sl Conservation} to bring up the context menu and select {\sl Add New Row}. A dialogue box will appear asking for {\sl Label for annotation}. Enter "Iron binding site" and click {\sl OK}. A new, empty, row appears.
+\exstep{Load the alignment at \textsf{http://www.jalview.org/tutorial/alignment.fa}. Right-click on the annotation label for {\sl Conservation} to bring up the context menu and select {\sl Add New Row}. A dialogue box will appear asking for {\sl Label for annotation}. Enter ``Iron binding site" and click {\sl OK}. A new, empty, row appears.
}
\exstep{
-Navigate to column 97. Select column 97 on the new annotation row. Right click on the selection and select {\sl Label} from the context menu. Enter "Fe" in the box and click {\sl OK}. Right-click on the selection again and select {\sl Colour}. Choose a colour from the colour chooser dialogue and click {\sl OK}. Press [ESC] to remove the selection.
+Navigate to column 97. Select column 97 on the new annotation row. Right click on the selection and select {\sl Label} from the context menu. Enter ``Fe" in the box and click {\sl OK}. Right-click on the selection again and select {\sl Colour}. Choose a colour from the colour chooser dialogue and click {\sl OK}. Press [ESC] to remove the selection.
}
-\exstep{ Select columns 70-77 on the annotation row. Right-click and choose {\sl Sheet} from the context menu. You will be prompted for a label. Enter "B" and press OK. A new line showing the sheet as an arrow appears. The colour of the label can be changed but not the colour of the sheet arrow.
+\exstep{ Select columns 70-77 on the annotation row. Right-click and choose {\sl Sheet} from the context menu. You will be prompted for a label. Enter ``B" and press OK. A new line showing the 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 annotation row that you just created. Select {\sl Export Annotation} and, in the {\bf Export Annotation} dialog box that will open, select the Jalview format and click the [To Textbox] button.
\exstep{Open the exported annotation in a text editor, and use the {\bf 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.}
}
-\exstep{ \label{viewannotfileex} Recover or recreate the secondary structure prediction that you made in exercise \ref{secstrpredex}. Use the {\sl File $\Rightarrow$ Export Annotation} function to view the jnet secondary structure prediction annotation row. Note the {\bf SEQUENCE\_REF} statements surrounding the row specifying the sequence association for the annotation. }
-}
+\label{viewannotfileex}\exstep{Recover or recreate the secondary structure
+prediction that you made in exercise \ref{secstrpredex}. Use the {\sl File $\Rightarrow$ Export Annotation} function to view the jnet secondary structure prediction annotation row. Note the {\bf SEQUENCE\_REF} statements surrounding the row specifying the sequence association for the annotation. } }
\section{Importing features from databases}
-
-Jalview supports feature retrieval from public databases either directly or {\sl via} the Distributed Annotation System (DAS\footnote{http://www.biodas.org/}). It includes built in parsers for Uniprot and EMBL records retrieved from the EBI. Sequences retrieved from these sources using the sequence fetcher (see section \ref{fetchseq}) will already posess features.
+\label{featuresfromdb}
+Jalview supports feature retrieval from public databases either directly or {\sl via} the Distributed Annotation System (DAS\footnote{http://www.biodas.org/}). It includes built in parsers for Uniprot and EMBL records retrieved from the EBI. Sequences retrieved from these sources using the sequence fetcher (see Section \ref{fetchseq}) will already posess features.
\subsection{Sequence Database Reference Retrieval}
\label{fetchdbrefs}
-Jalview maintains a list of external database references for each sequence in an alignment. These are listed in a tooltip when the mouse is moved over the sequence ID. Sequences retrieved using the sequence fetcher will always have at least one database reference, but alignments imported from a file, or retrieved from alignment datasources like PFAM, generally have no database references.
+Jalview maintains a list of external database references for each sequence in
+an alignment. These are listed in a tooltip when the mouse is moved over the
+sequence ID when the {\sl View $\Rightarrow$ Sequence ID Tooltip $\Rightarrow$
+Show Database Refs } option is enabled. Sequences retrieved using the sequence
+fetcher will always have at least one database reference, but alignments
+imported from an alignment file generally have no database references.
\subsubsection{Database References and Sequence Coordinate Systems}
-Jalview displays features in the local sequence's coordinate system which is given by its 'start' and 'end'. Any sequence features on the sequence will be rendered relative to the sequence's start position. If the start/end positions do not match the coordinate system from which the features were defined, then the features will be displayed incorrectly.
-
-\subsubsection{Automatic retrieval of a sequence's database references}
-Jalview includes a function to automatically verify and update each sequence's start and end numbering against any of the sequence databases that the {\sl Sequence Fetcher} has access to. This function is accessed by selecting {\sl Webservices $\Rightarrow$ Fetch DB References} from the Alignment window. When 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 matched against the local sequence, and if the local sequence is found to be a sub-sequence of the retrieved sequence then the local sequence's start/end numbering is updated. A new database reference mapping is created mapping the local sequence to the external database, and the local sequence inherits any additional annotation retrieved from the database sequence.
-
-The database retrieval process terminates when a valid mapping is found for a sequence, or if all database queries failed to retrieve a matching sequence.
+Jalview displays features in the local sequence's coordinate system which is
+given by its `start' and `end'. Any sequence features on the sequence will be
+rendered relative to the sequence's start position. If the start/end positions
+do not match the coordinate system from which the features were defined, then
+the features will be displayed incorrectly.
+
+\subsubsection{Automatically discovering a sequence's database references}
+Jalview includes a function to automatically verify and update each sequence's
+start and end numbering against any of the sequence databases that the {\sl
+Sequence Fetcher} has access to. This function is accessed from the {\sl
+Webservices $\Rightarrow$ Fetch DB References} sub-menu in the Alignment
+window. This menu allows you to query either the set of {\sl Standard
+Databases}, which includes EMBL, Uniprot, the PDB, and the currently selected
+DAS sequence sources, or just a specific datasource from one of the submenus.
+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
+matched against the local sequence, and if the local sequence is found to be a
+sub-sequence of the retrieved sequence then the local sequence's start/end
+numbering is updated. A new database reference mapping is created, mapping the
+local sequence to the external database, and the local sequence inherits any
+additional annotation retrieved from the database sequence.
+
+The database retrieval process terminates when a valid mapping is found for a
+sequence, or if all database queries failed to retrieve a matching sequence.
+Termination is indicated by the disappearance of the moving progress indicator
+on the alignment window. A dialog box may be shown once it completes which
+lists sequences for which records were found, but the sequence retrieved from
+the database did not exactly contain the sequence given in the alignment (the
+{\sl ``Sequence not 100\% match'' dialog box}).
\exercise{Retrieving Database References}{
\exstep{Load the example alignment at http://www.jalview.org/tutorial/alignment.fa}
-\exstep{Move the mouse over the sequence IDs to verify there are no database references for the sequences.}
+\exstep{Verify that there are no database references for the sequences by first
+checking that the {\sl View $\Rightarrow$ Sequence ID Tooltip $\Rightarrow$ Show
+Database IDs} option is selected, and then mousing over each sequence's ID.}
\exstep{Use the {\sl Webservices $\Rightarrow$ Fetch DB References} menu option to retrieve database IDs for the sequences.}
\exstep{Examine the tooltips for each sequence in the alignment as the retrieval progresses - note the appearance of new database references.}
+\exstep{Once the process has finished, save the alignment as a Jalview Project.
+
+Now close all the windows and open the project again, and verify that the database references and sequence features are still present on the alignment}
+
}
\subsection{Retrieving Features {\sl via} DAS}
-Jalview includes a client to retrieve features from DAS annotation servers. To retrieve features, select {\sl View $\Rightarrow$ Feature Settings\ldots} from the alignment window menu. Select the {\sl DAS Settings} tab in the Feature Settings Window (Figure \ref{das}). A list of DAS sources compiled from the currently configured DAS registry\footnote{By default, this will be the major public DAS server registry maintained by the Sanger Institute: http://www.dasregistry.org} is shown in the left hand pane. Highlighting an entry on the left brings up information about that source in the right hand panel.
+\label{dasfretrieval}
+Jalview includes a client to retrieve features from DAS annotation servers. To
+retrieve features, select {\sl View $\Rightarrow$ Feature Settings\ldots} from the alignment window menu. Select the {\sl DAS Settings} tab in the Feature Settings Window (Figure \ref{das}). A list of DAS sources compiled from the currently configured DAS registry\footnote{By default, this will be the major public DAS server registry maintained by the Sanger Institute: http://www.dasregistry.org} is shown in the left hand pane. Highlighting an entry on the left brings up information about that source in the right hand panel.
\begin{figure}[htbp]
\begin{center}
\end{center}
\end{figure}
+Select appropriate DAS sources as required then click on {\sl Fetch DAS
+Features}. If you know of additional sources not listed in the configured
+registry, then you may add them with the {\sl Add Local Source} button. Use
+the {\sl Authority},{\sl Type}, and {\sl Label} filters to restrict the list
+of sources to just those that will return features for the sequences in the
+alignment.
-
-Select appropriate DAS sources as required then click on {\sl Fetch DAS Features}. If you know of additional sources not listed in the by the configured registry, then you may add them with the {\sl Add Local Source} button. Use the {\sl Authority},{\sl Type}, and {\sl Label} filters to restrict the list of sources to just those that will return features for the sequences in the alignment.
-
-Following DAS feature retrieval, the {\sl Feature Settings} panel takes on a slightly different appearance (Figure \ref{das} (right)). Each data source is listed and groups of features from one data source can be selected/deselected by checking the labeled box at the top of the panel.
+Following DAS feature retrieval, the {\sl Feature Settings} panel takes on a
+slightly different appearance (Figure \ref{das} (right)). Each data source is
+listed and groups of features from one data source can be selected/deselected
+by checking the labeled box at the top of the panel.
\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 Services $\Rightarrow$ Fetch Database References } function described in section \ref{fetchdbrefs}. If a sequence is verified, then the start/end numbering will be adjusted to match the Uniprot record to ensure that features retrieved from the DAS source are rendered at the correct position.
+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 Services $\Rightarrow$ Fetch Database References } function described in Section \ref{fetchdbrefs}. If a sequence is verified, then the start/end numbering will be adjusted to match the Uniprot record to ensure that features retrieved from the DAS source are rendered at the correct position.
\subsubsection{Rate of feature retrieval}
Feature retrieval can take some time if a large number of sources is selected and if the alignment contains a large number of sequences. This is because Jalview only queries a particular DAS source with one sequence at a time, to avoid overloading it. As features are retrieved, they are immediately added to the current alignment view. The retrieved features are shown on the sequence and can be customised as described previously.
\exercise{Retrieving features with DAS}{
-\exstep{Load the alignment at \textsf{http://www.jalview.org/tutorial/alignment.fa}. Select {\sl View $\Rightarrow$ Sequence Features\ldots} from the alignment window menu. Select the {\sl DAS Settings} tab. A long list of available DAS sources is listed. Select a small number, eg Uniprot, DSSP, signalP and netoglyc. Click {\sl OK}. A window may prompt whether you wish Jalview to map the sequence IDs onto Uniprot IDs. Click {\sl Yes}. Jalview will start retrieving features. As features become available they will be mapped onto the alignment.
-}
-\exstep{If Jalview is taking too long to retrieve features, the process can be cancelled with the {\sl Cancel Fetch} button. Rolling the mouse cursor over the sequences reveals a large number of features annotated in the tool tip. Close the Feature Settings window.
-}
+\label{dasfeatretrexcercise}
+\exstep{Load the alignment at
+\textsf{http://www.jalview.org/tutorial/alignment.fa}. Select {\sl View
+$\Rightarrow$ Feature Settings \ldots} from the alignment window menu. Select
+the {\sl DAS Settings} tab. A long list of available DAS sources is listed. Select a small number, eg Uniprot, DSSP, signalP and netoglyc. Click {\sl OK}. A window may prompt whether you wish Jalview to map the sequence IDs onto Uniprot IDs. Click {\sl Yes}. Jalview will start retrieving features. As features become available they will be mapped onto the alignment. } \exstep{If Jalview is taking too long to retrieve features, the process can be cancelled with the {\sl Cancel Fetch} button. Rolling the mouse cursor over the sequences reveals a large number of features annotated in the tool tip. Close the Feature Settings window. }
\exstep{Move the mouse over the sequence ID panel. Non-positional features such as literature references and protein localisation predictions are given in the tooltip, below any database cross references associated with the sequence.}
\exstep{Search through the alignment to find a feature with a link symbol next to it. Right click to bring up the alignment view popup menu, and find a corresponding entry in the {\sl Link } sub menu. }
% TODO this doesn't work ! \includegraphics[width=.3in]{images/link.pdf}
\exstep{
-Select {\sl View $\Rightarrow$ Feature Settings\ldots} to reopen the Feature Settings window. All the loaded feature types should now be displayed. Those at the top of the list sit on top of and obscure those below. Move the feature settings window so that the alignment is visible and uncheck some of the feature types by clicking the tick box in the display column.. Observe how the alignment display changes. Note that unselected feature types do not appear in the tool tip.
+Select {\sl View $\Rightarrow$ Feature Settings\ldots} to reopen the Feature Settings window. All the loaded feature types should now be displayed. Those at the top of the list sit on top of and obscure those below. Move the feature settings window so that the alignment is visible and uncheck some of the feature types by clicking the tick box in the display column. Observe how the alignment display changes. Note that unselected feature types do not appear in the tool tip.
}
\exstep{Reorder the features by dragging feature types up and down the order in the Feature Settings panel. e.g. Click on {\sl CHAIN} then move the mouse downwards to drag it below {\sl DOMAIN}. Note that {\sl DOMAIN} is now shown on top of {\sl CHAIN} in the alignment window. Drag {\sl METAL} to the top of the list. Observe how the cysteine residues are now highlighted as they have a {\sl METAL} feature associated with them.
}
% TODO: describe working with features files and GFF
}
}
-\subsection{Sequence Feature File Formats}
-
-Jalview supports both GFF and its own Jalview Features file format for the import of sequence annotation. Features and alignment annotation are also extracted from other formats such as Stockholm, and AMSA. URL links may also be attached to features. See the online documentation for more details of the capabilities of the jalview features file.
-
-\section{Working with structures}
-Jalview provides integration between protein sequences and protein structures by providing a linked view of structures associated with protein sequences in the alignment. The Java based GPL molecular viewer Jmol\footnote{see the Jmol homepage www.jmol.org for more information.} is incorporated, enabling sophisticated molecular visualization to be prepared and investigated alongside analysis of associated sequences. \footnote{Earlier versions of Jalview included MCView - a simple main chain structure viewer. Structures are visualised as an alpha carbon trace and can be viewed, rotated and coloured in a structure viewer and the results interpreted on a sequence alignment.}
-
-PDB format files can be imported directly or structures can be retrieved from the the Macromolecular Structure Database (MSD) using the {\sl Sequence Fetcher}.
-
-\subsection{Automatic association of PDB structures with sequences}
-Jalview can automatically determine which structures are associated with a sequence, if that sequence has an ID from a public database such as Uniprot. Right-click on any sequence ID and select {\sl $<$Sequence ID$>$ $\Rightarrow$ Associate Structure with Sequence $\Rightarrow$ Discover PDB IDs } from the context menu (where {\sl $<$Sequence ID$>$} is the ID of the sequence on which you clicked) (Figure \ref{auto}). Jalview will attempt to associate the sequence with a Uniprot sequence and from there discover any associated PDB structures. This takes a few seconds and applies to all sequences in the alignment which have valid Uniprot IDs. On moving the cursor over the sequence ID the tool tip now shows the Uniprot ID and any associated PDB structures.
-
-\begin{figure}[htbp]
-\begin{center}
-%TODO fix formatting
-\parbox{1.5in}{
-{\centering
-\begin{center}
-\includegraphics[width=1.5in]{images/auto1.pdf}
-\end{center}}
-} \parbox{3.25in}{
-{\centering
-\begin{center}
-\includegraphics[scale=0.5]{images/auto2.pdf}
-\end{center}
-}
-} \parbox{1.5in}{
-{\centering
-\begin{center}
-\includegraphics[width=1.5in]{images/auto3.pdf}
-\end{center}
-}
-}
-
-\caption{{\bf Automatic PDB ID discovery.} The tooltip (left) indicates that no PDB structure has been associated with the sequence. After PDB ID discovery (center) the tool tip now indicates the Uniprot ID and any associated PDB structures (right)}
-\label{auto}
-\end{center}
-\end{figure}
-
-\subsection{Viewing Protein Structures}
-The structure viewer can be launched through the sequence ID context menu. Select {\sl Structure $\Rightarrow$ View PDB entry $\Rightarrow$ $<$PDB ID$>$}. The structure 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 (if available). Moving the mouse cursor over the sequence to which the structure is linked in the alignment panel highlights the respective residue by showing its sidechain. Moving the cursor over the structure shows a tooltip and highlights the corresponding residue in the alignment.
-
-\begin{figure}[htbp]
-\begin{center}
-\parbox{3in}{
-{\centering
-\begin{center}
-\includegraphics[scale=0.5]{images/structure1.pdf}
-\end{center}
-}
-}
-\parbox{3.2in}{
-{\centering
-\begin{center}
-\includegraphics[width=3in]{images/structure2.pdf}
-\end{center}
-}
-}
-\caption{{\bf Structure visualisation} The structure viewer is launched from the sequence ID context menu (left) and allows the structure to be visualised using the embedded Jmol molecular viewer (right). }
-\label{structure}
-\end{center}
-\end{figure}
-
-\subsubsection{Customising structure display}
-
-Structure display can be modified using the {\sl Colour} and {\sl View} menus in the structure viewer. The background colour can be modified by selecting the {\sl Colours $\Rightarrow$ Background Colour\ldots} option.
-
-By default, the structure will be coloured in the same way as the sequence in the associated alignment view. The structure can be coloured independently of the sequence by selecting an appropriate colour scheme from the {\sl Colours} menu. It can be coloured according to the alignment using the {\sl Colours $\Rightarrow$ By Sequence } option. The image in the structure viewer can be output to EPS or PNG format via the {\sl File $\Rightarrow$ Save As $\Rightarrow$ \ldots} submenu. The mapping between the structure and the sequence (How well and which parts of the structure relate to the sequence) can be viewed with the {\sl File $\Rightarrow$ View Mapping} menu option.
-
-\subsubsection{Using the Jmol visualization interface }
-
-Jmol has a comprehensive set of selection and visualization functions that are accessed from the Jmol popup menu (by right-clicking in the Jmol window or by clicking the Jmol logo). Molecule colour and rendering style can be manipulated, and distance measurements and molecular surfaces can be added to the view. It also has its own Rasmol\footnote{see www.rasmol.org}-like scripting language, and this is used by Jalview to store the state of a Jmol visualization within Jalview archives, in addition to the PDB data file originally loaded or retrieved by Jalview. To access the Jmol scripting environment directly, use the {\sl Jmol $\Rightarrow$ Console} menu option.
-
-
-\exercise{Viewing Structures}{
-\exstep{Load the alignment at \textsf{http://www.jalview.org/examples/exampleFile.jar}. Right-click on the sequence ID label for any of the sequences (e.g. {\sl FER1\_SPIOL}) to bring up the context menu. Select {\sl FER1\_SPIOL $\Rightarrow$ Associate Structure with Sequence $\Rightarrow$ Discover PDB ids}. Jalview will now attempt to find PDB structures for the sequences in the alignment.
-}
-\exstep{
-Right-click on the sequence ID for {\sl FER1\_SPIOL}. Select { \sl FER1\_SPIOL $\Rightarrow$ View PDB Entry: 1A70} A structure viewing window appears. Rotate the molecule by clicking and dragging in the structure viewing box. Zoom with the mouse scroll wheel.
-}
-\exstep{Roll the mouse cursor along the {\sl FER1\_SPIOL} sequence in the alignment. Note that if a residue in the sequence maps to one in the structure, a label will appear next to that residue in the structure viewer. Move the mouse over the structure. Placing the mouse over a part of the structure will bring up a tool tip indicating the name and number of that residue. The corresponding residue in the sequence is highlighted in black. Clicking the alpha carbon toggles the highlight and residue label on and off. Try this by clicking on a set of three or four adjacent residues so that the labels are persistent, then finding where they are in the sequence.
-}
-\exstep{Select {\sl Colours $\Rightarrow$ Background Colour\ldots} from the structure viewer menu and choose a suitable colour. Press {\sl OK} to apply this. Select {\sl File $\Rightarrow$ Save As $\Rightarrow$ PNG} and save the image. View this with your web browser.
-}
-\exstep{Select {\sl File $\Rightarrow$ View Mapping} from the structure viewer menu. A new window opens showing the residue by residue alignment between the sequence and the structure.}
-\exstep{Select {\sl File $\Rightarrow$ Save $\Rightarrow$ PDB file} and choose a new filename to save the PDB file. Once the file is saved, open the location in your file browser (or explorer window) and drag the PDB file that you just saved on to the Jalview desktop (or load it from the {\sl Jalview Desktop $\Rightarrow$ Input Alignment $\Rightarrow$ From File } menu). Verify that you can open and view the associated structure from the sequence ID pop-up menu's {\sl Structure } submenu in the new alignment window.}
-
-\exstep{Right click on the structure to bring up the Jmol window. Explore the menu options. Try to change the style of molecular display - by first using the {\sl Jmol $\Rightarrow$ Select $\Rightarrow$ all} command, and then the {\sl Jmol $\Rightarrow$ Style $\Rightarrow$ Scheme $\Rightarrow$ Ball and stick} command.}
-\exstep{Use the {\sl File $\Rightarrow$ Save As .. } function to save the alignment as a Jalview Project. Now close the alignment and the structure view, and load the project file you just saved.
-
-Verify that the Jmol display is as it was when you just saved the file.}
+\subsection{Colouring features by score or description
+text}
+\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 feature type's popup menu, which is opened by
+right-clicking the feature type's color in the 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'
+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.
+
+Graduated feature colour schemes can also be used 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 by 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.
+
+\subsection{Using features to re-order the alignment}
+\label{featureordering}
+The presence of sequence features on certain sequences or in a particular
+region of an alignment can quantitatively identify important trends in
+the aligned sequences. In this case, it is more useful to
+re-order the alignment based on the number of features or their associated scores, rather than simply re-colour the aligned sequences. The sequence feature settings
+dialog box provides two buttons: `Seq sort by Density' and `Seq sort by
+Score', that allow you to reorder the alignment according to the number of
+sequence features present on each sequence, and also according to any scores
+associated with a feature. Each of these buttons uses the currently displayed
+features to determine the ordering, but
+if you wish to re-order the alignment using a single type of feature, then you can do this from the feature type's
+popup menu. Simply right-click the type's style in the Feature Settings dialog
+box, and select one of the {\sl Sort by score} and {\sl Sort by density}
+options to re-order the alignment. Finally, if a specific region is selected,
+then only features found in that region of the alignment will be used to
+create the new alignment ordering.
+
+\exercise{Shading and sorting alignments using sequence features}{
+\label{shadingorderingfeatsex}
+\exstep{Re-load the alignment from \ref{dasfeatretrexcercise}.
+}
+\exstep{Open the
+feature settings panel, and, after first clearing the current
+selection, press the {\em Seq Sort by Density} button a few times.}
+\exstep{Use the DAS fetcher to retrieve the Kyte and Doolittle Hydrophobicity
+scores for the protein sequences in the alignment.
+{\sl Hint: the nickname for the das source is `KD$\_$hydrophobicity'.}}
+\exstep{Change the feature settings so only the hydrophobicity features are
+displayed. Mouse over the annotation and also export and examine the GFF and
+Jalview features file to better understand how the hydrophobicity measurements
+are recorded.}
+\exstep{Apply a graduated colourscheme to the hydrophobicity annotation to
+reveal the variation in average hydrophobicity across the alignment.}
+\exstep{Select a range of alignment columns, and use one of the sort by feature buttons to order the alignment according to that region's average
+hydrophobicity.}
+\exstep{Save the alignment as a project, for use in exercise
+\ref{threshgradfeaturesex}.} }
+
+\exercise{Shading alignments with combinations of graduated feature
+colourschemes}{
+\label{threshgradfeaturesex}
+\exstep{Reusing the annotated alignment from exercise
+\ref{shadingorderingfeatsex}, experiment with the colourscheme threshold to
+highlight the most, or least hydrophobic regions. Note how the colour scheme icon for the feature type changes when you change the threshold type.}
+\exstep{Change the colourscheme so
+that features at the threshold are always coloured grey, and the most
+hydrophobic residues are coloured red, regardless of the threshold value
+({\em hint - there is a switch on the dialog to do this for you}).}
+\exstep{Enable the Uniprot {\em chain} annotation in the feature settings
+display and re-order the features so it is visible under the hydrophobicity
+annotation.}
+\exstep{Apply a graduated colourscheme to the {\em chain}
+annotation so that it distinguishes the different canonical names associated
+with the mature polypeptide chains.}
+\exstep{Export the alignment's sequence features using the Jalview sequence
+feature file format, to see how the different types of graduated feature
+colour styles are encoded. }
}
-
\section{Working with DNA}
\label{workingwithnuc}
-Jalview was originally developed for the analysis of protein sequences, but now includes some specific features for working with nucleic acid sequences and alignments. Nucleotide sequences and alignments are recognised based on the presence the nucleotide symbols [ACGT] of greater than 85\% in the sequences. Built in codon-translation tables can be used to translate ORFs into peptides for further analysis. EMBL 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 nucleotide sequence. Mappings are used to to transfer annotation between nucleic acid and protein sequences, and to dynamically highlight regions in one sequence that correspond to the position of the mouse pointer in another.
-
+Jalview was originally developed for the analysis of protein sequences, but
+now includes some specific features for working with nucleic acid sequences
+and alignments. Nucleotide sequences and alignments are recognised 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 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
+nucleotide sequence. Mappings are used to to transfer annotation between
+nucleic acid and protein sequences, and to dynamically highlight regions in
+one sequence that correspond to the position of the mouse pointer in another.
+%TODO Working with Nucleic acid sequences and structures.
\subsection{Alignment and Colouring}
-Jalview provides a simple colourscheme for DNA bases, but does not apply any specific conservation or substitution score model for the shading of nucleotide alignments. However, pairwise alignments performed using the {\sl Alignment Window $\Rightarrow$ Calculations $\Rightarrow$ Pairwise Alignment \ldots} option will utilise an identity score matrix to calculate alignment score when alignment two nucleotide sequences.
+Jalview provides a simple colourscheme for DNA bases, but does not apply any
+specific conservation or substitution score model for the shading of
+nucleotide alignments. However, pairwise alignments performed using the {\sl
+Alignment Window $\Rightarrow$ Calculations $\Rightarrow$ Pairwise Alignment
+\ldots} option will utilise an identity score matrix to calculate alignment
+score when aligning two nucleotide sequences.
\subsubsection{Aligning Nucleic Acid Sequences}
-Jalview only has limited knowledge of the capabilities of the programs that are made available to it {\sl via} web services. In particular, only the ClustalW and MAFFT programs will successfuly recognise and align nucleic acid sequences. MAFFT will also choose an appropriate parameter model. Whilst Muscle may appear to align DNA, it simply treats the base symbols as amino-acids, often leading to a poor quality alignment. Furthermore, it will almost certainly fail to align RNA containing Uracil bases, since 'U' is not a valid one-letter amino acid code.
+Jalview only has limited knowledge of the capabilities of the programs that
+are made available to it {\sl via} web services, so it is up to you, the user,
+to decide which service to use when working with nucleic acid sequences. The
+table below shows which alignment programs are appropriate for nucleotide alignment.
+
+\begin{table}{}
+\centering
+\begin{tabular}{|l|c|l|}
+Program& NA support& Notes\\
+\hline
+ClustalW& Yes&\begin{minipage}[f]{3in}
+Default is to autodetect nucleotide
+sequences. Editable parameters include nucleotide substitution matrices and
+distance metrics.
+\end{minipage}
+\\
+\hline
+Muscle& Yes (treat U as T)&\begin{minipage}[f]{3in}
+Default is to autodetect nucleotide
+sequences. Editable parameters include nucleotide substitution matrices and
+distance metrics.
+\end{minipage}
+\\
+\hline
+MAFFT& Yes&\begin{minipage}[f]{3in}
+Will autodetect nucleotide sequences and use a hardwired substitution model
+(all amino-acid sequence related parameters are ignored). Unknown whether
+substitution model treats Uracil specially.
+\end{minipage}
+\\
+\hline
+ProbCons& No&\begin{minipage}[f]{3in}
+ProbCons has no special support for aligning nucleotide sequences. Whilst an
+alignment will be returned, it is unlikely to be reliable.
+\end{minipage}
+\\
+\hline
+T-COFFEE& Yes&\begin{minipage}[f]{3in}
+Sequence type is automatically detected and an appropriate
+parameter set used as required. A range of nucleotide specific
+score models are available.\end{minipage}
+\\
+\end{tabular}
+\caption{{\bf JABAWS Alignment programs suitable for aligning nucleic acid
+sequences.} All JABAWS alignment services will return an alignment if provided
+with RNA or DNA sequences, with varying reliability. No programs include
+support for taking RNA secondary structure prediction into account when
+aligning sequences.}
+\label{nucleomsatools}
+\end{table}
\subsection{Translate cDNA}
-The {\sl Calculations $\Leftarrow$ Translate cDNA} function in the alignment window is only available when working with a nucleic acid alignment. It uses the standard codon translation table given in the online help to translate a nucleotide alignment, or the currently selected region, into a set of aligned peptide sequences. Any features or annotation present on the nucleotide alignment will also be translated, allowing DNA alignment analysis results to be transferred on to peptide products for further investigation.
+The {\sl Calculations $\Rightarrow$ Translate cDNA} function in the alignment
+window is only available when working with a nucleic acid alignment. It uses the standard codon translation table given in the online help to translate a nucleotide alignment, or the currently selected region, into a set of aligned peptide sequences. Any features or annotation present on the nucleotide alignment will also be translated, allowing DNA alignment analysis results to be transferred on to peptide products for further investigation.
\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 and extracted 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 and extracted 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.
}\parbox{3in}{
\begin{center}
%\begin{figure}[htbp]
\subsection{Coding regions from EMBL records}
-Many EMBL records that can be retrieved from the sequence fetcher contain exons. Coding regions will be marked as features on the EMBL 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 Uniprot sequence's coordinate system with the coding regions annotated on the EMBL sequence. Jalview utilises cross-reference information in two ways.
+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
+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
+Uniprot sequence's coordinate system with the coding regions annotated on the
+EMBL sequence. Jalview utilises cross-reference information in two ways.
\subsubsection{Retrieval of Protein or DNA Cross References}
The {\sl Calculations $\Rightarrow$ 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 product(s).
\label{dnadasfeatures}
\includegraphics[width=5in]{images/dnadasfeatures.pdf}
-\caption{Uniprot and PDB sum features retrieved {\sl via} DAS and mapped onto coding regions of EMBL record V00488}
+\caption{Uniprot and PDB sum features retrieved {\sl via} DAS and mapped onto
+coding regions of EMBL record V00488 (an earlier version of Jalview is shown
+here).}
\end{center}
\end{figure}
% \section{Customising Jalview}
% \subsection{Setting preferences}
%
-% The Jalview Desktop stores configuration and history information in a file stored in the users home directory, called '.jalview\_properties'. Many of the options stored in this file are presented in the {\sl Desktop $\Rightarrow$ Tools $\Rightarrow$ Preferences\ldots} dialog. These preferences include default settings for : {\bf Visual} layout settings for alignment views and controlling the display of the default alignment, {\bf Connection} preferences such as the standard set of URL paths that are available from the links menu and the URL which is opened when a Sequence's ID is double clicked, {\bf Editing} settings like {\sl Pad Gaps} and autocalculation of consensus, {\bf Output} settings control the degree of meta-information written in alignment files and mode of EPS figure generation, and finally the {\bf Das Settings} which allows the default DAS sources to be configured.
+% The Jalview Desktop stores configuration and history information in a file stored in the users home directory, called `.jalview\_properties'. Many of the options stored in this file are presented in the {\sl Desktop $\Rightarrow$ Tools $\Rightarrow$ Preferences\ldots} dialog. These preferences include default settings for : {\bf Visual} layout settings for alignment views and controlling the display of the default alignment, {\bf Connection} preferences such as the standard set of URL paths that are available from the links menu and the URL which is opened when a Sequence's ID is double clicked, {\bf Editing} settings like {\sl Pad Gaps} and autocalculation of consensus, {\bf Output} settings control the degree of meta-information written in alignment files and mode of EPS Figuregeneration, and finally the {\bf Das Settings} which allows the default DAS sources to be configured.
%
% \subsection{Adding your own URL links}
%
git://source.jalview.org / jalview-manual.git / blobdiff