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Clustalw.pm
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Clustalw.pm
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# $Id$
#
# BioPerl module for Bio::Tools::Run::Alignment::Clustalw
#
# Cared for by
#
# Copyright Peter Schattner
#
# You may distribute this module under the same terms as perl itself
# POD documentation - main docs before the code
=head1 NAME
Bio::Tools::Run::Alignment::Clustalw - Object for the calculation of a
multiple sequence alignment from a set of unaligned sequences or
alignments using the Clustalw program
=head1 SYNOPSIS
# Build a clustalw alignment factory
@params = ('ktuple' => 2, 'matrix' => 'BLOSUM');
$factory = Bio::Tools::Run::Alignment::Clustalw->new(@params);
# Pass the factory a list of sequences to be aligned.
$inputfilename = 't/data/cysprot.fa';
$aln = $factory->align($inputfilename); # $aln is a SimpleAlign object.
# or
$seq_array_ref = \@seq_array;
# where @seq_array is an array of Bio::Seq objects
$aln = $factory->align($seq_array_ref);
# Or one can pass the factory a pair of (sub)alignments
#to be aligned against each other, e.g.:
$aln = $factory->profile_align($aln1,$aln2);
# where $aln1 and $aln2 are Bio::SimpleAlign objects.
# Or one can pass the factory an alignment and one or more unaligned
# sequences to be added to the alignment. For example:
$aln = $factory->profile_align($aln1,$seq); # $seq is a Bio::Seq object.
There are various additional options and input formats available. See
the DESCRIPTION section that follows for additional details.
=head1 DESCRIPTION
Note: this DESCRIPTION only documents the (Bio)perl interface to
Clustalw. Clustalw, itself, is a large & complex program - for more
information regarding clustalw, please see the clustalw documentation
which accompanies the clustalw distribution. Clustalw is available
from (among others) ftp://ftp.ebi.ac.uk/pub/software/. Clustalw.pm has
been tested so far only under Linux. I expect that it should also work
under other Unix systems. However, since the module is currently
implemented using (unix) system calls, extensive modification may be
necessary before Clustalw.pm would work under non-Unix operating
systems (eg Windows, MacOS). Clustalw.pm has only been tested using
version 1.8 of clustalw. Compatibility with earlier versions of the
clustalw program is currently unknown. Before running Clustalw.pm
successfully it will be necessary: to install clustalw on your system,
to edit the variable $clustdir in Clustalw.pm to point to the clustalw
program, and to ensure that users have execute privilieges for the
clustalw program.
Bio::Tools::Run::Alignment::Clustalw.pm: is an object for performing a
multiple sequence alignment from a set of unaligned sequences and/or
sub-alignments by means of the clustalw program.
Initially, a clustalw "factory object" is created. Optionally, the
factory may be passed most of the parameters or switches of the
clustalw program, e.g.:
@params = ('ktuple' => 2, 'matrix' => 'BLOSUM');
$factory = Bio::Tools::Run::Alignment::Clustalw->new(@params);
Any parameters not explicitly set will remain as the defaults of the
clustalw program. Additional parameters and switches (not available
in clustalw) may also be set. Currently, the only such parameter is
"quiet", which when set to a non-zero value, suppresses clustalw
terminal output. Not all clustalw parameters are supported at this
stage.
By default, Clustalw.pm output is returned solely in a the form of a
BioPerl Bio::SimpleAlign object which can then be printed and/or saved
in multiple formats using the AlignIO.pm module. Optionally the raw
clustalw output file can be saved if the calling script specifies an
output file (with the clustalw parameter OUTFILE). Currently only the
GCG-MSF output file formats is supported.
Other parameters and features (such as those corresponding to tree
production) have not been implemented yet in Perl format.
Alignment parameters can be changed and/or examined at any time after
the factory has been created. The program checks that any
parameter/switch being set/read is valid. However, currently no
additional checks are included to check that parameters are of the
proper type (eg string or numeric) or that their values are within the
proper range. As an example, to change the value of the clustalw
parameter ktuple to 3 and subsequently to check its value one would
write:
$ktuple = 3;
$factory->ktuple($ktuple);
$get_ktuple = $factory->ktuple();
Once the factory has been created and the appropriate parameters set,
one can call the method align() to align a set of unaligned sequences,
or call profile_align() to add one or more sequences or a second
alignment to an initial alignment.
Input to align() may consist of a set of unaligned sequences in the
form of the name of file containing the sequences. For example,
$inputfilename = 't/data/cysprot.fa'; $aln =
$factory-E<gt>align($inputfilename);
Alternately one can create an array of Bio::Seq objects somehow
$str = Bio::SeqIO->new(-file=> 't/data/cysprot.fa', '-format' => 'Fasta');
@seq_array =();
while ( my $seq = $str->next_seq() ) {push (@seq_array, $seq) ;}
and pass the factory a reference to that array
$seq_array_ref = \@seq_array;
$aln = $factory->align($seq_array_ref);
In either case, align() returns a reference to a SimpleAlign object
which can then be displayed, stored, or converted to a UnivAlign
object for further manipulation.
Once an initial alignment exists, one can pass the factory additional
sequence(s) to be added (ie aligned) to the original alignment. The
alignment can be passed as either an alignment file or a
Bio:SimpleAlign object. The unaligned sequence(s) can be passed as a
filename or as an array of BioPerl sequence objects or as a single
BioPerl Seq object. For example (to add a single sequence to an
alignment),
$str = Bio::AlignIO->new(-file=> 't/data/cysprot1a.msf');
$aln = $str->next_aln();
$str1 = Bio::SeqIO->new(-file=> 't/data/cysprot1b.fa');
$seq = $str1->next_seq();
$aln = $factory->profile_align($aln,$seq);
In either case, profile_align() returns a reference to a SimpleAlign
object containing a new SimpleAlign object of the alignment with the
additional sequence(s) added in.
Finally one can pass the factory a pair of (sub)alignments to be
aligned against each other. The alignments can be passed in the form
of either a pair of alignment files or a pair of Bio:SimpleAlign
objects. For example,
$profile1 = 't/data/cysprot1a.msf';
$profile2 = 't/data/cysprot1b.msf';
$aln = $factory->profile_align($profile1,$profile2);
or
$str1 = Bio::AlignIO->new(-file=> 't/data/cysprot1a.msf');
$aln1 = $str1->next_aln();
$str2 = Bio::AlignIO->new(-file=> 't/data/cysprot1b.msf');
$aln2 = $str2->next_aln();
$aln = $factory->profile_align($aln1,$aln2);
In either case, profile_align() returns a reference to a SimpleAlign
object containing an (super)alignment of the two input alignments.
For more examples of syntax and use of Clustalw.pm, the user is
encouraged to run the script Clustalw.t is the bioperl/t directory.
Note: Clustalw.pm is still under development. Various features of the
clustalw program have not yet been implemented. If you would like
that a specific clustalw feature be added to this perl interface, let
me know.
These can be specified as paramters when instantiating a new TCoffee
object, or through get/set methods of the same name (lowercase).
=head1 PARAMETER FOR ALIGNMENT COMPUTATION
=head2 KTUPLE
Title : KTUPLE
Description : (optional) set the word size to be used in the alignment
This is the size of exactly matching fragment that is used.
INCREASE for speed (max= 2 for proteins; 4 for DNA),
DECREASE for sensitivity.
For longer sequences (e.g. >1000 residues) you may
need to increase the default
=head2 TOPDIAGS
Title : TOPDIAGS
Description : (optional) number of best diagonals to use
The number of k-tuple matches on each diagonal
(in an imaginary dot-matrix plot) is calculated.
Only the best ones (with most matches) are used in
the alignment. This parameter specifies how many.
Decrease for speed; increase for sensitivity.
=head2 WINDOW
Title : WINDOW
Description : (optional) window size
This is the number of diagonals around each of the 'best'
diagonals that will be used. Decrease for speed;
increase for sensitivity.
=head2 PAIRGAP
Title : PAIRGAP
Description : (optional) gap penalty for pairwise alignments
This is a penalty for each gap in the fast alignments.
It has little affect on the speed or sensitivity except
for extreme values.
=head2 FIXEDGAP
Title : FIXEDGAP
Description : (optional) fixed length gap penalty
=head2 FLOATGAP
Title : FLOATGAP
Description : (optional) variable length gap penalty
=head2 MATRIX
Title : MATRIX
Default : PAM100 for DNA - PAM250 for protein alignment
Description : (optional) substitution matrix used in the multiple
alignments. Depends on the version of clustalw as to
what default matrix will be used
PROTEIN WEIGHT MATRIX leads to a new menu where you are
offered a choice of weight matrices. The default for
proteins in version 1.8 is the PAM series derived by
Gonnet and colleagues. Note, a series is used! The
actual matrix that is used depends on how similar the
sequences to be aligned at this alignment step
are. Different matrices work differently at each
evolutionary distance.
DNA WEIGHT MATRIX leads to a new menu where a single
matrix (not a series) can be selected. The default is
the matrix used by BESTFIT for comparison of nucleic
acid sequences.
=head2 TYPE
Title : TYPE
Description : (optional) sequence type: protein or DNA. This allows
you to explicitly overide the programs attempt at
guessing the type of the sequence. It is only useful
if you are using sequences with a VERY strange
composition.
=head2 OUTPUT
Title : OUTPUT
Description : (optional) clustalw supports GCG or PHYLIP or PIR or
Clustal format. See the Bio::AlignIO modules for
which formats are supported by bioperl.
=head2 OUTFILE
Title : OUTFILE
Description : (optional) Name of clustalw output file. If not set
module will erase output file. In any case alignment will
be returned in the form of SimpleAlign objects
=head2 TRANSMIT
Title : TRANSMIT
Description : (optional) transitions not weighted. The default is to
weight transitions as more favourable than other
mismatches in DNA alignments. This switch makes all
nucleotide mismatches equally weighted.
=head1 FEEDBACK
=head2 Mailing Lists
User feedback is an integral part of the evolution of this and other
Bioperl modules. Send your comments and suggestions preferably to one
of the Bioperl mailing lists. Your participation is much appreciated.
bioperl-l@bioperl.org - General discussion
http://bio.perl.org/MailList.html - About the mailing lists
=head2 Reporting Bugs
Report bugs to the Bioperl bug tracking system to help us keep track
the bugs and their resolution. Bug reports can be submitted via
email or the web:
bioperl-bugs@bio.perl.org
http://bio.perl.org/bioperl-bugs/
=head1 AUTHOR - Peter Schattner
Email schattner@alum.mit.edu
=head1 APPENDIX
The rest of the documentation details each of the object
methods. Internal methods are usually preceded with a _
=cut
#'
package Bio::Tools::Run::Alignment::Clustalw;
use vars qw($AUTOLOAD @ISA $PROGRAM $PROGRAMDIR
$TMPDIR $TMPOUTFILE @CLUSTALW_SWITCHES @CLUSTALW_PARAMS
@OTHER_SWITCHES %OK_FIELD);
use strict;
use Bio::Seq;
use Bio::SeqIO;
use Bio::SimpleAlign;
use Bio::AlignIO;
use Bio::Root::Root;
use Bio::Root::IO;
use Bio::Factory::ApplicationFactoryI;
@ISA = qw(Bio::Root::Root Bio::Root::IO Bio::Factory::ApplicationFactoryI);
# You will need to enable Clustalw to find the clustalw program. This
# can be done in (at least) two ways:
#
# 1. define an environmental variable CLUSTALDIR:
# export CLUSTALDIR=/home/peter/clustalw1.8
#
# 2. include a definition of an environmental variable CLUSTALDIR in
# every script that will use Clustal.pm.
# $ENV{CLUSTALDIR} = '/home/peter/clustalw1.8/';
BEGIN {
if (defined $ENV{CLUSTALDIR}) {
$PROGRAMDIR = $ENV{CLUSTALDIR} || '';
$PROGRAM = Bio::Root::IO->catfile($PROGRAMDIR,
'clustalw'.($^O =~ /mswin/i ?'.exe':''));
}
else {
$PROGRAM = 'clustalw';
}
@CLUSTALW_PARAMS = qw(KTUPLE TOPDIAGS WINDOW PAIRGAP FIXEDGAP
FLOATGAP MATRIX TYPE TRANSIT DNAMATRIX OUTFILE
GAPOPEN GAPEXT MAXDIV GAPDIST HGAPRESIDUES PWMATRIX
PWDNAMATRIX PWGAPOPEN PWGAPEXT SCORE TRANSWEIGHT
SEED HELIXGAP OUTORDER STRANDGAP LOOPGAP TERMINALGAP
HELIXENDIN HELIXENDOUT STRANDENDIN STRANDENDOUT PROGRAM);
@CLUSTALW_SWITCHES = qw(HELP CHECK OPTIONS NEGATIVE NOWEIGHTS ENDGAPS
NOPGAP NOHGAP NOVGAP KIMURA TOSSGAPS);
@OTHER_SWITCHES = qw(QUIET);
# Authorize attribute fields
foreach my $attr ( @CLUSTALW_PARAMS, @CLUSTALW_SWITCHES,
@OTHER_SWITCHES ) { $OK_FIELD{$attr}++; }
}
sub new {
my ($class,@args) = @_;
my $self = $class->SUPER::new(@args);
# to facilitiate tempfile cleanup
$self->_initialize_io();
my ($attr, $value);
(undef,$TMPDIR) = $self->tempdir(CLEANUP=>1);
(undef,$TMPOUTFILE) = $self->tempfile(-dir => $TMPDIR);
while (@args) {
$attr = shift @args;
$value = shift @args;
next if( $attr =~ /^-/ ); # don't want named parameters
if ($attr eq 'PROGRAM') {
$self->program($value);
next;
}
$self->$attr($value);
}
if (! defined $self->program) {
$self->program($PROGRAM);
}
unless ($self->exists_clustal()) {
if( $self->verbose >= 0 ) {
warn "Clustalw program not found as ".$self->program." or not executable. \n Clustalw can be obtained from eg- http://corba.ebi.ac.uk/Biocatalog/Alignment_Search_software.html/ \n";
}
}
return $self;
}
sub AUTOLOAD {
my $self = shift;
my $attr = $AUTOLOAD;
$attr =~ s/.*:://;
$attr = uc $attr;
$self->throw("Unallowed parameter: $attr !") unless $OK_FIELD{$attr};
$self->{$attr} = shift if @_;
return $self->{$attr};
}
=head2 exists_clustal()
Title : exists_clustal
Usage : $clustalfound = Bio::Tools::Run::Alignment::Clustalw->exists_clustal()
Function: Determine whether clustalw program can be found on current host
Example :
Returns : 1 if clustalw program found at expected location, 0 otherwise.
Args : none
=cut
sub exists_clustal {
my $self = shift;
return Bio::Root::IO->exists_exe($self->program);
}
=head2 program
Title : program
Usage : $obj->program($newval)
Function:
Returns : value of program
Args : newvalue (optional)
=cut
sub program{
my $self = shift;
if( @_ ) {
my $value = shift;
$self->{'program'} = $value;
}
return $self->{'program'};
}
=head2 version
Title : version
Usage : exit if $prog->version() < 1.8
Function: Determine the version number of the program
Example :
Returns : float or undef
Args : none
=cut
sub version {
my ($self) = @_;
return undef unless $self->exists_clustal;
my $string = `clustalw -- ` ;
$string =~ /\(([\d.]+)\)/;
return $1 || undef;
}
=head2 align
Title : align
Usage :
$inputfilename = 't/data/cysprot.fa';
$aln = $factory->align($inputfilename);
or
$seq_array_ref = \@seq_array; @seq_array is array of Seq objs
$aln = $factory->align($seq_array_ref);
Function: Perform a multiple sequence alignment
Example :
Returns : Reference to a SimpleAlign object containing the
sequence alignment.
Args : Name of a file containing a set of unaligned fasta sequences
or else an array of references to Bio::Seq objects.
Throws an exception if argument is not either a string (eg a
filename) or a reference to an array of Bio::Seq objects. If
argument is string, throws exception if file corresponding to string
name can not be found. If argument is Bio::Seq array, throws
exception if less than two sequence objects are in array.
=cut
sub align {
my ($self,$input) = @_;
my ($temp,$infilename, $seq);
my ($attr, $value, $switch);
# Create input file pointer
$infilename = $self->_setinput($input);
if (!$infilename) {$self->throw("Bad input data (sequences need an id ) or less than 2 sequences in $input !");}
# Create parameter string to pass to clustalw program
my $param_string = $self->_setparams();
# run clustalw
my $aln = $self->_run('align', $infilename,$param_string);
}
#################################################
=head2 profile_align
Title : profile_align
Usage :
Function: Perform an alignment of 2 (sub)alignments
Example :
Returns : Reference to a SimpleAlign object containing the (super)alignment.
Args : Names of 2 files containing the subalignments
or references to 2 Bio::SimpleAlign objects.
Throws an exception if arguments are not either strings (eg filenames)
or references to SimpleAlign objects.
=cut
sub profile_align {
my ($self,$input1,$input2) = @_;
my ($temp,$infilename1,$infilename2,$input,$seq);
# Create input file pointer
$infilename1 = $self->_setinput($input1,1);
$infilename2 = $self->_setinput($input2,2);
if (!$infilename1 || !$infilename2) {$self->throw("Bad input data: $input1 or $input2 !");}
unless ( -e $infilename1 and -e $infilename2) {$self->throw("Bad input file: $input1 or $input2 !");}
# Create parameter string to pass to clustalw program
my $param_string = $self->_setparams();
# run clustalw
my $aln = $self->_run('profile-aln', $infilename1,
$infilename2, $param_string);
}
#################################################
=head2 _run
Title : _run
Usage : Internal function, not to be called directly
Function: makes actual system call to clustalw program
Example :
Returns : nothing; clustalw output is written to a
temporary file $TMPOUTFILE
Args : Name of a file containing a set of unaligned fasta sequences
and hash of parameters to be passed to clustalw
=cut
sub _run {
my ($self,$command,$infile1,$infile2,$param_string) = @_;
my $instring;
if ($command =~ /align/) {
$instring = "-infile=$infile1";
$param_string .= " $infile2";
}
if ($command =~ /profile/) {
$instring = "-profile1=$infile1 -profile2=$infile2";
chmod 0777, $infile1,$infile2;
$command = '-profile';
}
$self->debug( "Program ".$self->program."\n");
my $commandstring = $self->program." $command"." $instring".
" -output=gcg". " $param_string";
$self->debug( "clustal command = $commandstring");
my $status = system($commandstring);
$self->throw( "Clustalw call ($commandstring) crashed: $? \n") unless $status==0;
my $outfile = $self->outfile() || $TMPOUTFILE ;
# retrieve alignment (Note: MSF format for AlignIO = GCG format of clustalw)
my $in = Bio::AlignIO->new(-file => $outfile, '-format' => 'MSF');
my $aln = $in->next_aln();
# Clean up the temporary files created along the way...
# Replace file suffix with dnd to find name of dendrogram file(s) to delete
foreach my $f ( $infile1, $infile2 ) {
$f =~ s/\.[^\.]*$// ;
unlink $f .'.dnd' if( $f ne '' );
}
return $aln;
}
=head2 _setinput()
Title : _setinput
Usage : Internal function, not to be called directly
Function: Create input file for clustalw program
Example :
Returns : name of file containing clustalw data input
Args : Seq or Align object reference or input file name
=cut
sub _setinput {
my ($self, $input, $suffix) = @_;
my ($infilename, $seq, $temp, $tfh);
# suffix is used to distinguish alignment files If $input is not a
# reference it better be the name of a file with the sequence/
# alignment data...
unless (ref $input) {
# check that file exists or throw
$infilename = $input;
unless (-e $input) {return 0;}
return $infilename;
}
# $input may be an array of BioSeq objects...
if (ref($input) eq "ARRAY") {
# Open temporary file for both reading & writing of BioSeq array
($tfh,$infilename) = $self->tempfile(-dir=>$TMPDIR);
$temp = Bio::SeqIO->new('-fh'=>$tfh,
'-format' =>'Fasta');
# Need at least 2 seqs for alignment
unless (scalar(@$input) > 1) {return 0;}
foreach $seq (@$input) {
unless (defined $seq &&
$seq->isa("Bio::PrimarySeqI") and $seq->id() )
{return 0;}
$temp->write_seq($seq);
}
$temp->close();
return $infilename;
}
# $input may be a SimpleAlign object.
elsif (ref($input) eq "Bio::SimpleAlign") {
# Open temporary file for both reading & writing of SimpleAlign object
if ($suffix ==1 || $suffix== 2 ) {
($tfh,$infilename) = $self->tempfile(-dir=>$TMPDIR);
}
$temp = Bio::AlignIO->new('-fh'=> $tfh,
'-format' => 'Fasta');
$temp->write_aln($input);
return $infilename;
}
# or $input may be a single BioSeq object (to be added to a previous alignment)
elsif (ref($input) && $input->isa("Bio::PrimarySeqI") && $suffix==2) {
# Open temporary file for both reading & writing of BioSeq object
($tfh,$infilename) = $self->tempfile();
$temp = Bio::SeqIO->new(-fh=> $tfh, '-format' =>'Fasta');
$temp->write_seq($input);
return $infilename;
}
return 0;
}
=head2 _setparams()
Title : _setparams
Usage : Internal function, not to be called directly
Function: Create parameter inputs for clustalw program
Example :
Returns : parameter string to be passed to clustalw
during align or profile_align
Args : name of calling object
=cut
sub _setparams {
my ($attr, $value, $self);
$self = shift;
my $param_string = "";
for $attr ( @CLUSTALW_PARAMS ) {
$value = $self->$attr();
next unless (defined $value);
my $attr_key = lc $attr; #put params in format expected by clustalw
$attr_key = ' -'.$attr_key;
$param_string .= $attr_key.'='.$value;
}
for $attr ( @CLUSTALW_SWITCHES) {
$value = $self->$attr();
next unless ($value);
my $attr_key = lc $attr; #put switches in format expected by clustalw
$attr_key = ' -'.$attr_key;
$param_string .= $attr_key ;
# $attr_key = '-'.$attr_key;
# $param_string .= '"'.$attr_key.'",';
}
# Set default output file if no explicit output file selected
unless ($param_string =~ /outfile/) {
$param_string .= " -outfile=$TMPOUTFILE" ;
}
if ($self->quiet() || $self->verbose() < 0) {
$param_string .= ' >/dev/null';
}
return $param_string;
}
1; # Needed to keep compiler happy