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#
# BioPerl module for Bio::Tools::Mdust
#
# Please direct questions and support issues to <bioperl-l@bioperl.org>
#
# Cared for by Donald Jackson, donald.jackson@bms.com
#
# Copyright Donald Jackson
#
# You may distribute this module under the same terms as perl itself
# POD documentation - main docs before the code
=head1 NAME
Mdust - Perl extension for Mdust nucleotide filtering
=head1 SYNOPSIS
use Bio::Tools::Run::Mdust;
my $mdust = Bio::Tools::Run::Mdust->new();
$mdust->run($bio_seq_object);
=head1 DESCRIPTION
Perl wrapper for the nucleic acid complexity filtering program
B<mdust> as available from TIGR via
L<http://www.tigr.org/tdb/tgi/software/>. Takes a Bio::SeqI or
Bio::PrimarySeqI object of type DNA as input.
If a Bio::Seq::RichSeqI is passed then the low-complexity regions will
be added to the feature table of the target object as
Bio::SeqFeature::Generic items with primary tag = 'Excluded' .
Otherwise a new target object will be returned with low-complexity
regions masked (by N's or other character as specified by maskchar()).
The mdust executable must be in a directory specified with either the
PATH or MDUSTDIR environment variable.
=head1 SEE ALSO
L<mdust>,
L<Bio::PrimarySeq>,
L<Bio::Seq::RichSeq>,
L<Bio::SeqFeature::Generic>
=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
the Bioperl mailing list. Your participation is much appreciated.
bioperl-l@bioperl.org - General discussion
http://bioperl.org/wiki/Mailing_lists - About the mailing lists
=head2 Support
Please direct usage questions or support issues to the mailing list:
I<bioperl-l@bioperl.org>
rather than to the module maintainer directly. Many experienced and
reponsive experts will be able look at the problem and quickly
address it. Please include a thorough description of the problem
with code and data examples if at all possible.
=head2 Reporting Bugs
Report bugs to the Bioperl bug tracking system to help us keep track
of the bugs and their resolution. Bug reports can be submitted via
the web:
=head1 AUTHOR
Donald Jackson (donald.jackson@bms.com)
=head1 APPENDIX
The rest of the documentation details each of the object methods.
Internal methods are usually preceded with a _
=cut
require 5.005_62;
use strict;
use vars qw($AUTOLOAD $PROGRAMNAME @ARGNAMES @MASKCHARS $VERSION @ISA);
@ISA = qw(Bio::Root::Root Bio::Tools::Run::WrapperBase);
@ARGNAMES = qw(TARGET WSIZE CUTOFF MASKCHAR COORDS TMPDIR DEBUG);
$PROGRAMNAME = 'mdust';
@MASKCHARS = qw(N X L);
=head2 new
Title : new
Usage : my $mdust = Bio::Tools::Run::Mdust->new( -target => $target_bioseq)
Purpose : Create a new mdust object
Returns : A Bio::Seq object
Args : target - Bio::Seq object for masking - alphabet MUST be DNA.
wsize - word size for masking (default = 3)
cutoff - cutoff score for masking (default = 28)
maskchar - character for replacing masked regions (default = N)
coords - boolean - indicate low-complexity regions as
Bio::SeqFeature::Generic
objects with primary tag 'Excluded',
do not change sequence (default 0)
tmpdir - directory for storing temporary files
debug - boolean - toggle debugging output,
do not remove temporary files
Notes : All of the arguments can also be get/set with their own accessors, such as:
my $wsize = $mdust->wsize();
When processing multiple sequences, call Bio::Tools::Run::Mdust->new() once
then pass each sequence as an argument to the target() or run() methods.
=cut
sub new {
my ($proto, @args) = @_;
my $pkg = ref($proto) || $proto;
my %args;
my $self = { wsize => undef,
cutoff => undef,
maskchar => undef,
coords => 0,
};
bless ($self, $pkg);
@args{@ARGNAMES} = $self->_rearrange(\@ARGNAMES, @args);
# load target first since it requires special handling
$self->target($args{'TARGET'}) if ($args{'TARGET'});
# package settings
$self->{'coords'} = $args{'COORDS'} if (defined $args{'COORDS'});
$self->{'tmpdir'} = $args{'TMPDIR'} || $ENV{'TMPDIR'} || $ENV{'TMP'} || '.';
# mdust options
$self->{'wsize'} = $args{'WSIZE'} if (defined $args{'WSIZE'});
$self->{'cutoff'} = $args{'CUTOFF'} if (defined $args{'CUTOFF'});
$self->{'maskchar'} = $args{'MASKCHAR'} if (defined $args{'CUTOFF'});
# set debugging
$self->verbose($args{'DEBUG'});
return $self;
}
=head2 run
Title : run
Usage : $mdust->run();
Purpose : Run mdust on the target sequence
Args : target (optional) - Bio::Seq object of alphabet DNA for masking
Returns : Bio::Seq object with masked sequence or low-complexity regions added to feature table.
=cut
sub run {
my ($self, $target) = @_;
if ($target) {
$self->target($target);
}
return $self->_run_mdust;
}
sub program_dir {
return Bio::Root::IO->catfile($ENV{MDUSTDIR}) if $ENV{MDUSTDIR};
}
sub program_name {
return $PROGRAMNAME;
}
sub _run_mdust {
# open a pipe to the mdust command. Pass in sequence(s?) as fasta
# files on STDIN, recover filtered seqs on STDOUT
my ($self) = @_;
my $target = $self->target or warn "No target sequence specified\n" && return undef;
# make sure program is available - doesn't seem to check
#my $executable = $self->executable('mdust', 1);
# add options
my $mdust_cmd = $self->program_path;
$mdust_cmd .= " -w " . $self->wsize if (defined $self->wsize);
$mdust_cmd .= " -v " . $self->cutoff if (defined $self->cutoff);
$mdust_cmd .= " -m " . $self->maskchar if (defined $self->maskchar);
$mdust_cmd .= " -c" if ($self->coords);
print STDERR "Running mdust: $mdust_cmd\n" if ($self->debug);
my $maskedfile = $self->_maskedfile;
eval {
my $pid = open (MDUST, "| $mdust_cmd > $maskedfile"); # bind STDIN of mdust to filehandle
local $| = 1;
my $seqout = Bio::SeqIO->new(-fh => \*MDUST, -format => 'Fasta');
$seqout->write_seq($target);
close MDUST; # need to do this to get output to flush!
};
$self->throw($@) if ($@);
my $rval;
if ($self->coords) {
$self->_parse_coords($maskedfile);
$rval = $self->target;
}
else { # replace original seq w/ masked seq
my $seqin = Bio::SeqIO->new(-file=>$maskedfile, -format => 'Fasta');
$rval = $seqin->next_seq
}
unlink $maskedfile unless $self->save_tempfiles;
return $rval;
}
=head2 target
Title : target
Usage : $mdust->target($bio_seq)
Purpose : Set/get the target (sequence to be filtered).
Returns : Target Bio::Seq object
Args : Bio::SeqI or Bio::PrimarySeqI object using the DNA alphabet (optional)
Note : If coordinate parsing is selected ($mdust->coords = 1) then target
MUST be a Bio::Seq::RichSeqI object. Passing a RichSeqI object automatically
turns on coordinate parsing.
=cut
sub target {
my ($self, $targobj) = @_;
if ($targobj) {
return $self->_set_target($targobj);
}
else {
return $self->{'target'};
}
}
sub _set_target {
my ($self, $targobj) = @_;
unless ($targobj->isa('Bio::SeqI') or ($targobj->isa('Bio::PrimarySeqI'))) {
$self->throw( -text => "Target must be passed as a Bio::SeqI or Bio::PrimarySeqI object",
-class => 'Bio::Root::BadParameter',
-value => $targobj );
}
if ($self->coords) {
unless ($targobj->isa('Bio::Seq::RichSeqI')) {
$self->throw( -text => "Target must be passed as a Bio::Seq::RichSeqSeqI object when coords == 1",
-class => 'Bio::Root::BadParameter',
-value => $targobj );
}
}
elsif ($targobj->isa('Bio::Seq::RichSeqI')) {
$self->coords(1);
}
unless ($targobj->alphabet eq 'dna') {
$self->throw( -text => "Target must be a DNA sequence",
-class => 'Bio::Root::BadParameter',
-value => $targobj );
}
$self->{'target'} = $targobj;
return 1;
}
sub _maskedfile {
my ($self, $file) = @_;
my $tmpdir = $self->tempdir;
if ($file) {
$self->{'maskedfile'} = $file;
# add some sanity chex for writability?
}
elsif (!$self->{'maskedfile'}) {
($self->{'maskedfh'},$self->{'maskedfile'}) =
$self->io->tempfile(-dir=>$self->tempdir());
}
return $self->{'maskedfile'};
}
sub _parse_coords {
my ($self, $file) = @_;
my $target = $self->target;
open(FILE, $file) or die "Unable to open $file: $!";
while (<FILE>) {
chomp;
s/\r//;
my ($seq, $length, $mstart, $mstop) = split(/\t/);
# add masked region as a SeqFeature in target
my $masked = Bio::SeqFeature::Generic->new( -start => $mstart,
-end => $mstop,
);
$masked->primary_tag('Excluded');
$masked->source_tag('mdust');
$target->add_SeqFeature($masked);
}
return 1;
}
=head2 maskchar
Title : maskchar
Usage : $mdust->maskchar('N')
Purpose : Set/get the character for masking low-complexity regions
Returns : True on success
Args : Either N (default), X or L (lower case)
=cut
sub maskchar {
my ($self, $maskchar) = @_;
return $self->{'maskchar'} unless (defined $maskchar);
unless ( grep {$maskchar eq $_} @MASKCHARS ) {
$self->throw( -text => "maskchar must be one of N, X or L",
-class => 'Bio::Root::BadParameter',
-value => $maskchar );
}
$self->{'maskchar'} = $maskchar;
1;
}
sub DESTROY {
my $self= shift;
unless ( $self->save_tempfiles ) {
$self->cleanup();
}
$self->SUPER::DESTROY();
}
sub AUTOLOAD {
my ($self, $value) = @_;
my $name = $AUTOLOAD;
$name =~ s/.+:://;
return if ($name eq 'DESTROY');
if (defined $value) {
$self->{$name} = $value;
}
unless (exists $self->{$name}) {
warn "Attribute $name not defined for ", ref($self), "\n" if ($self->debug);
return undef;
}
return $self->{$name};
}
1;