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# BioPerl module for Bio::Tools::Mdust |
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# |
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# Please direct questions and support issues to |
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# |
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# Cared for by Donald Jackson, donald.jackson@bms.com |
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# |
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# Copyright Donald Jackson |
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# |
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# You may distribute this module under the same terms as perl itself |
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# POD documentation - main docs before the code |
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=head1 NAME |
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Mdust - Perl extension for Mdust nucleotide filtering |
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=head1 SYNOPSIS |
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use Bio::Tools::Run::Mdust; |
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my $mdust = Bio::Tools::Run::Mdust->new(); |
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$mdust->run($bio_seq_object); |
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=head1 DESCRIPTION |
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Perl wrapper for the nucleic acid complexity filtering program |
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B as available from TIGR via |
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L. Takes a Bio::SeqI or |
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Bio::PrimarySeqI object of type DNA as input. |
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If a Bio::Seq::RichSeqI is passed then the low-complexity regions will |
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be added to the feature table of the target object as |
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Bio::SeqFeature::Generic items with primary tag = 'Excluded' . |
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Otherwise a new target object will be returned with low-complexity |
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regions masked (by N's or other character as specified by maskchar()). |
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The mdust executable must be in a directory specified with either the |
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PATH or MDUSTDIR environment variable. |
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=head1 SEE ALSO |
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L, |
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L, |
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L, |
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L |
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=head1 FEEDBACK |
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=head2 Mailing Lists |
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User feedback is an integral part of the evolution of this and other |
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Bioperl modules. Send your comments and suggestions preferably to |
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the Bioperl mailing list. Your participation is much appreciated. |
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bioperl-l@bioperl.org - General discussion |
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http://bioperl.org/wiki/Mailing_lists - About the mailing lists |
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=head2 Support |
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Please direct usage questions or support issues to the mailing list: |
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I |
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rather than to the module maintainer directly. Many experienced and |
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reponsive experts will be able look at the problem and quickly |
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address it. Please include a thorough description of the problem |
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with code and data examples if at all possible. |
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=head2 Reporting Bugs |
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Report bugs to the Bioperl bug tracking system to help us keep track |
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of the bugs and their resolution. Bug reports can be submitted via |
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the web: |
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http://redmine.open-bio.org/projects/bioperl/ |
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=head1 AUTHOR |
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Donald Jackson (donald.jackson@bms.com) |
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=head1 APPENDIX |
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The rest of the documentation details each of the object methods. |
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Internal methods are usually preceded with a _ |
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=cut |
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package Bio::Tools::Run::Mdust; |
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require 5.005_62; |
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use strict; |
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use Bio::SeqIO; |
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use Bio::SeqFeature::Generic; |
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use Bio::Root::Root; |
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use Bio::Root::IO; |
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use Bio::Tools::Run::WrapperBase; |
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use vars qw($AUTOLOAD $PROGRAMNAME @ARGNAMES @MASKCHARS $VERSION @ISA); |
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@ISA = qw(Bio::Root::Root Bio::Tools::Run::WrapperBase); |
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@ARGNAMES = qw(TARGET WSIZE CUTOFF MASKCHAR COORDS TMPDIR DEBUG); |
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$PROGRAMNAME = 'mdust'; |
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@MASKCHARS = qw(N X L); |
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=head2 new |
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Title : new |
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Usage : my $mdust = Bio::Tools::Run::Mdust->new( -target => $target_bioseq) |
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Purpose : Create a new mdust object |
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Returns : A Bio::Seq object |
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Args : target - Bio::Seq object for masking - alphabet MUST be DNA. |
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wsize - word size for masking (default = 3) |
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cutoff - cutoff score for masking (default = 28) |
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maskchar - character for replacing masked regions (default = N) |
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coords - boolean - indicate low-complexity regions as |
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Bio::SeqFeature::Generic |
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objects with primary tag 'Excluded', |
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do not change sequence (default 0) |
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tmpdir - directory for storing temporary files |
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debug - boolean - toggle debugging output, |
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do not remove temporary files |
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Notes : All of the arguments can also be get/set with their own accessors, such as: |
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my $wsize = $mdust->wsize(); |
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When processing multiple sequences, call Bio::Tools::Run::Mdust->new() once |
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then pass each sequence as an argument to the target() or run() methods. |
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=cut |
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sub new { |
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my ($proto, @args) = @_; |
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my $pkg = ref($proto) || $proto; |
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my %args; |
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my $self = { wsize => undef, |
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cutoff => undef, |
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maskchar => undef, |
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coords => 0, |
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}; |
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bless ($self, $pkg); |
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@args{@ARGNAMES} = $self->_rearrange(\@ARGNAMES, @args); |
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# load target first since it requires special handling |
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$self->target($args{'TARGET'}) if ($args{'TARGET'}); |
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# package settings |
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$self->{'coords'} = $args{'COORDS'} if (defined $args{'COORDS'}); |
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$self->{'tmpdir'} = $args{'TMPDIR'} || $ENV{'TMPDIR'} || $ENV{'TMP'} || '.'; |
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# mdust options |
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$self->{'wsize'} = $args{'WSIZE'} if (defined $args{'WSIZE'}); |
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$self->{'cutoff'} = $args{'CUTOFF'} if (defined $args{'CUTOFF'}); |
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$self->{'maskchar'} = $args{'MASKCHAR'} if (defined $args{'CUTOFF'}); |
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# set debugging |
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$self->verbose($args{'DEBUG'}); |
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return $self; |
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} |
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=head2 run |
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Title : run |
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Usage : $mdust->run(); |
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Purpose : Run mdust on the target sequence |
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Args : target (optional) - Bio::Seq object of alphabet DNA for masking |
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Returns : Bio::Seq object with masked sequence or low-complexity regions added to feature table. |
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=cut |
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sub run { |
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my ($self, $target) = @_; |
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if ($target) { |
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$self->target($target); |
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} |
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return $self->_run_mdust; |
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} |
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sub program_dir { |
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return Bio::Root::IO->catfile($ENV{MDUSTDIR}) if $ENV{MDUSTDIR}; |
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} |
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sub program_name { |
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return $PROGRAMNAME; |
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} |
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sub _run_mdust { |
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# open a pipe to the mdust command. Pass in sequence(s?) as fasta |
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# files on STDIN, recover filtered seqs on STDOUT |
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my ($self) = @_; |
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200
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my $target = $self->target or warn "No target sequence specified\n" && return undef; |
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# make sure program is available - doesn't seem to check |
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#my $executable = $self->executable('mdust', 1); |
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# add options |
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my $mdust_cmd = $self->program_path; |
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$mdust_cmd .= " -w " . $self->wsize if (defined $self->wsize); |
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$mdust_cmd .= " -v " . $self->cutoff if (defined $self->cutoff); |
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$mdust_cmd .= " -m " . $self->maskchar if (defined $self->maskchar); |
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$mdust_cmd .= " -c" if ($self->coords); |
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print STDERR "Running mdust: $mdust_cmd\n" if ($self->debug); |
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my $maskedfile = $self->_maskedfile; |
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eval { |
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my $pid = open (MDUST, "| $mdust_cmd > $maskedfile"); # bind STDIN of mdust to filehandle |
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216
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0
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0
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local $| = 1; |
217
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0
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0
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my $seqout = Bio::SeqIO->new(-fh => \*MDUST, -format => 'Fasta'); |
218
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0
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0
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$seqout->write_seq($target); |
219
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0
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0
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close MDUST; # need to do this to get output to flush! |
220
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}; |
221
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222
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0
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0
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0
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$self->throw($@) if ($@); |
223
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224
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0
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0
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my $rval; |
225
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226
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0
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0
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0
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if ($self->coords) { |
227
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0
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0
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$self->_parse_coords($maskedfile); |
228
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0
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0
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$rval = $self->target; |
229
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} |
230
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else { # replace original seq w/ masked seq |
231
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0
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0
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my $seqin = Bio::SeqIO->new(-file=>$maskedfile, -format => 'Fasta'); |
232
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0
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0
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$rval = $seqin->next_seq |
233
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} |
234
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235
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0
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0
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0
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unlink $maskedfile unless $self->save_tempfiles; |
236
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237
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0
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0
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return $rval; |
238
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239
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} |
240
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241
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=head2 target |
242
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243
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Title : target |
244
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Usage : $mdust->target($bio_seq) |
245
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Purpose : Set/get the target (sequence to be filtered). |
246
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Returns : Target Bio::Seq object |
247
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Args : Bio::SeqI or Bio::PrimarySeqI object using the DNA alphabet (optional) |
248
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Note : If coordinate parsing is selected ($mdust->coords = 1) then target |
249
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MUST be a Bio::Seq::RichSeqI object. Passing a RichSeqI object automatically |
250
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|
turns on coordinate parsing. |
251
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252
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|
=cut |
253
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254
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|
sub target { |
255
|
1
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1
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1
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3
|
my ($self, $targobj) = @_; |
256
|
|
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257
|
1
|
50
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|
3
|
if ($targobj) { |
258
|
1
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5
|
return $self->_set_target($targobj); |
259
|
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|
} |
260
|
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|
else { |
261
|
0
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|
0
|
return $self->{'target'}; |
262
|
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|
} |
263
|
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264
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|
} |
265
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266
|
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267
|
|
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|
|
sub _set_target { |
268
|
1
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|
|
1
|
|
3
|
my ($self, $targobj) = @_; |
269
|
|
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|
270
|
1
|
50
|
33
|
|
|
6
|
unless ($targobj->isa('Bio::SeqI') or ($targobj->isa('Bio::PrimarySeqI'))) { |
271
|
0
|
|
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|
|
0
|
$self->throw( -text => "Target must be passed as a Bio::SeqI or Bio::PrimarySeqI object", |
272
|
|
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|
|
-class => 'Bio::Root::BadParameter', |
273
|
|
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|
|
-value => $targobj ); |
274
|
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|
|
} |
275
|
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|
|
276
|
|
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|
|
|
|
277
|
1
|
50
|
|
|
|
9
|
if ($self->coords) { |
|
|
50
|
|
|
|
|
|
278
|
0
|
0
|
|
|
|
0
|
unless ($targobj->isa('Bio::Seq::RichSeqI')) { |
279
|
0
|
|
|
|
|
0
|
$self->throw( -text => "Target must be passed as a Bio::Seq::RichSeqSeqI object when coords == 1", |
280
|
|
|
|
|
|
|
-class => 'Bio::Root::BadParameter', |
281
|
|
|
|
|
|
|
-value => $targobj ); |
282
|
|
|
|
|
|
|
} |
283
|
|
|
|
|
|
|
} |
284
|
|
|
|
|
|
|
elsif ($targobj->isa('Bio::Seq::RichSeqI')) { |
285
|
1
|
|
|
|
|
4
|
$self->coords(1); |
286
|
|
|
|
|
|
|
} |
287
|
|
|
|
|
|
|
|
288
|
|
|
|
|
|
|
|
289
|
1
|
50
|
|
|
|
11
|
unless ($targobj->alphabet eq 'dna') { |
290
|
0
|
|
|
|
|
0
|
$self->throw( -text => "Target must be a DNA sequence", |
291
|
|
|
|
|
|
|
-class => 'Bio::Root::BadParameter', |
292
|
|
|
|
|
|
|
-value => $targobj ); |
293
|
|
|
|
|
|
|
} |
294
|
|
|
|
|
|
|
|
295
|
1
|
|
|
|
|
17
|
$self->{'target'} = $targobj; |
296
|
1
|
|
|
|
|
3
|
return 1; |
297
|
|
|
|
|
|
|
|
298
|
|
|
|
|
|
|
} |
299
|
|
|
|
|
|
|
|
300
|
|
|
|
|
|
|
sub _maskedfile { |
301
|
0
|
|
|
0
|
|
0
|
my ($self, $file) = @_; |
302
|
0
|
|
|
|
|
0
|
my $tmpdir = $self->tempdir; |
303
|
|
|
|
|
|
|
|
304
|
0
|
0
|
|
|
|
0
|
if ($file) { |
|
|
0
|
|
|
|
|
|
305
|
0
|
|
|
|
|
0
|
$self->{'maskedfile'} = $file; |
306
|
|
|
|
|
|
|
# add some sanity chex for writability? |
307
|
|
|
|
|
|
|
} |
308
|
|
|
|
|
|
|
elsif (!$self->{'maskedfile'}) { |
309
|
0
|
|
|
|
|
0
|
($self->{'maskedfh'},$self->{'maskedfile'}) = |
310
|
|
|
|
|
|
|
$self->io->tempfile(-dir=>$self->tempdir()); |
311
|
|
|
|
|
|
|
} |
312
|
0
|
|
|
|
|
0
|
return $self->{'maskedfile'}; |
313
|
|
|
|
|
|
|
|
314
|
|
|
|
|
|
|
} |
315
|
|
|
|
|
|
|
|
316
|
|
|
|
|
|
|
sub _parse_coords { |
317
|
0
|
|
|
0
|
|
0
|
my ($self, $file) = @_; |
318
|
0
|
|
|
|
|
0
|
my $target = $self->target; |
319
|
0
|
0
|
|
|
|
0
|
open(FILE, $file) or die "Unable to open $file: $!"; |
320
|
0
|
|
|
|
|
0
|
while () { |
321
|
0
|
|
|
|
|
0
|
chomp; |
322
|
0
|
|
|
|
|
0
|
s/\r//; |
323
|
0
|
|
|
|
|
0
|
my ($seq, $length, $mstart, $mstop) = split(/\t/); |
324
|
|
|
|
|
|
|
|
325
|
|
|
|
|
|
|
# add masked region as a SeqFeature in target |
326
|
0
|
|
|
|
|
0
|
my $masked = Bio::SeqFeature::Generic->new( -start => $mstart, |
327
|
|
|
|
|
|
|
-end => $mstop, |
328
|
|
|
|
|
|
|
); |
329
|
0
|
|
|
|
|
0
|
$masked->primary_tag('Excluded'); |
330
|
0
|
|
|
|
|
0
|
$masked->source_tag('mdust'); |
331
|
|
|
|
|
|
|
|
332
|
0
|
|
|
|
|
0
|
$target->add_SeqFeature($masked); |
333
|
|
|
|
|
|
|
} |
334
|
0
|
|
|
|
|
0
|
return 1; |
335
|
|
|
|
|
|
|
} |
336
|
|
|
|
|
|
|
|
337
|
|
|
|
|
|
|
=head2 maskchar |
338
|
|
|
|
|
|
|
|
339
|
|
|
|
|
|
|
Title : maskchar |
340
|
|
|
|
|
|
|
Usage : $mdust->maskchar('N') |
341
|
|
|
|
|
|
|
Purpose : Set/get the character for masking low-complexity regions |
342
|
|
|
|
|
|
|
Returns : True on success |
343
|
|
|
|
|
|
|
Args : Either N (default), X or L (lower case) |
344
|
|
|
|
|
|
|
|
345
|
|
|
|
|
|
|
=cut |
346
|
|
|
|
|
|
|
|
347
|
|
|
|
|
|
|
sub maskchar { |
348
|
0
|
|
|
0
|
1
|
0
|
my ($self, $maskchar) = @_; |
349
|
|
|
|
|
|
|
|
350
|
0
|
0
|
|
|
|
0
|
return $self->{'maskchar'} unless (defined $maskchar); |
351
|
|
|
|
|
|
|
|
352
|
0
|
0
|
|
|
|
0
|
unless ( grep {$maskchar eq $_} @MASKCHARS ) { |
|
0
|
|
|
|
|
0
|
|
353
|
0
|
|
|
|
|
0
|
$self->throw( -text => "maskchar must be one of N, X or L", |
354
|
|
|
|
|
|
|
-class => 'Bio::Root::BadParameter', |
355
|
|
|
|
|
|
|
-value => $maskchar ); |
356
|
|
|
|
|
|
|
} |
357
|
0
|
|
|
|
|
0
|
$self->{'maskchar'} = $maskchar; |
358
|
|
|
|
|
|
|
|
359
|
0
|
|
|
|
|
0
|
1; |
360
|
|
|
|
|
|
|
} |
361
|
|
|
|
|
|
|
|
362
|
|
|
|
|
|
|
sub DESTROY { |
363
|
1
|
|
|
1
|
|
1901
|
my $self= shift; |
364
|
1
|
50
|
|
|
|
24
|
unless ( $self->save_tempfiles ) { |
365
|
1
|
|
|
|
|
17
|
$self->cleanup(); |
366
|
|
|
|
|
|
|
} |
367
|
1
|
|
|
|
|
5
|
$self->SUPER::DESTROY(); |
368
|
|
|
|
|
|
|
} |
369
|
|
|
|
|
|
|
|
370
|
|
|
|
|
|
|
|
371
|
|
|
|
|
|
|
sub AUTOLOAD { |
372
|
2
|
|
|
2
|
|
5
|
my ($self, $value) = @_; |
373
|
2
|
|
|
|
|
3
|
my $name = $AUTOLOAD; |
374
|
2
|
|
|
|
|
9
|
$name =~ s/.+:://; |
375
|
|
|
|
|
|
|
|
376
|
2
|
50
|
|
|
|
7
|
return if ($name eq 'DESTROY'); |
377
|
|
|
|
|
|
|
|
378
|
|
|
|
|
|
|
|
379
|
2
|
100
|
|
|
|
4
|
if (defined $value) { |
380
|
1
|
|
|
|
|
3
|
$self->{$name} = $value; |
381
|
|
|
|
|
|
|
} |
382
|
|
|
|
|
|
|
|
383
|
2
|
50
|
|
|
|
25
|
unless (exists $self->{$name}) { |
384
|
0
|
0
|
|
|
|
0
|
warn "Attribute $name not defined for ", ref($self), "\n" if ($self->debug); |
385
|
0
|
|
|
|
|
0
|
return undef; |
386
|
|
|
|
|
|
|
} |
387
|
|
|
|
|
|
|
|
388
|
2
|
|
|
|
|
8
|
return $self->{$name}; |
389
|
|
|
|
|
|
|
} |
390
|
|
|
|
|
|
|
|
391
|
|
|
|
|
|
|
1; |