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# BioPerl module for Bio::Tools::Sim4::Results |
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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 Ewan Birney |
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# and Hilmar Lapp |
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# |
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# Copyright Ewan Birney and Hilmar Lapp |
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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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Bio::Tools::Sim4::Results - Results of one Sim4 run |
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=head1 SYNOPSIS |
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# to preset the order of EST and genomic file as given on the sim4 |
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# command line: |
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my $sim4 = Bio::Tools::Sim4::Results->new(-file => 'result.sim4', |
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-estfirst => 1); |
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# to let the order be determined automatically (by length comparison): |
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$sim4 = Bio::Tools::Sim4::Results->new( -file => 'sim4.results' ); |
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# filehandle: |
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$sim4 = Bio::Tools::Sim4::Results->new( -fh => \*INPUT ); |
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# parse the results |
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while(my $exonset = $sim4->next_exonset()) { |
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# $exonset is-a Bio::SeqFeature::Generic with Bio::Tools::Sim4::Exons |
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# as sub features |
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print "Delimited on sequence ", $exonset->seq_id(), |
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"from ", $exonset->start(), " to ", $exonset->end(), "\n"; |
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foreach my $exon ( $exonset->sub_SeqFeature() ) { |
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# $exon is-a Bio::SeqFeature::FeaturePair |
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print "Exon from ", $exon->start, " to ", $exon->end, |
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" on strand ", $exon->strand(), "\n"; |
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# you can get out what it matched using the est_hit attribute |
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my $homol = $exon->est_hit(); |
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print "Matched to sequence ", $homol->seq_id, |
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" at ", $homol->start," to ", $homol->end, "\n"; |
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} |
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} |
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# essential if you gave a filename at initialization (otherwise the file |
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# stays open) |
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$sim4->close(); |
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=head1 DESCRIPTION |
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The sim4 module provides a parser and results object for sim4 output. The |
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sim4 results are specialised types of SeqFeatures, meaning you can add them |
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to AnnSeq objects fine, and manipulate them in the "normal" seqfeature manner. |
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The sim4 Exon objects are Bio::SeqFeature::FeaturePair inherited objects. The |
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$esthit = $exon-Eest_hit() is the alignment as a feature on the matching |
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object (normally, an EST), in which the start/end points are where the hit |
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lies. |
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To make this module work sensibly you need to run |
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sim4 genomic.fasta est.database.fasta |
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or |
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sim4 est.fasta genomic.database.fasta |
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To get the sequence identifiers recorded for the first sequence, too, use |
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A=4 as output option for sim4. |
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One fiddle here is that there are only two real possibilities to the matching |
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criteria: either one sequence needs reversing or not. Because of this, it |
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is impossible to tell whether the match is in the forward or reverse strand |
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of the genomic DNA. We solve this here by assuming that the genomic DNA is |
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always forward. As a consequence, the strand attribute of the matching EST is |
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unknown, and the strand attribute of the genomic DNA (i.e., the Exon object) |
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will reflect the direction of the hit. |
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See the documentation of parse_next_alignment() for abilities of the parser |
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to deal with the different output format options of sim4. |
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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 one |
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of the Bioperl mailing lists. 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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the bugs and their resolution. Bug reports can be submitted via the |
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web: |
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https://github.com/bioperl/bioperl-live/issues |
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=head1 AUTHOR - Ewan Birney, Hilmar Lapp |
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Ewan Birney Ebirney-at-sanger.ac.ukE |
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Hilmar Lapp Ehlapp-at-gmx.netE or Ehilmar.lapp-at-pharma.novartis.comE. |
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=head1 APPENDIX |
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The rest of the documentation details each of the object methods. Internal methods are usually preceded with a _ |
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=cut |
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# Let the code begin... |
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package Bio::Tools::Sim4::Results; |
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use strict; |
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use File::Basename; |
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use Bio::Root::Root; |
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use Bio::Tools::Sim4::Exon; |
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use base qw(Bio::Tools::AnalysisResult); |
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sub _initialize_state { |
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my($self,@args) = @_; |
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# call the inherited method first |
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my $make = $self->SUPER::_initialize_state(@args); |
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my ($est_is_first) = $self->_rearrange([qw(ESTFIRST)], @args); |
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delete($self->{'_est_is_first'}); |
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$self->{'_est_is_first'} = $est_is_first if(defined($est_is_first)); |
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$self->analysis_method("Sim4"); |
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} |
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=head2 analysis_method |
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Usage : $sim4->analysis_method(); |
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Purpose : Inherited method. Overridden to ensure that the name matches |
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/sim4/i. |
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Returns : String |
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Argument : n/a |
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=cut |
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#------------- |
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sub analysis_method { |
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#------------- |
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my ($self, $method) = @_; |
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if($method && ($method !~ /sim4/i)) { |
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$self->throw("method $method not supported in " . ref($self)); |
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} |
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return $self->SUPER::analysis_method($method); |
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} |
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=head2 parse_next_alignment |
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Title : parse_next_alignment |
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Usage : @exons = $sim4_result->parse_next_alignment; |
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foreach $exon (@exons) { |
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# do something |
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} |
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Function: Parses the next alignment of the Sim4 result file and returns the |
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found exons as an array of Bio::Tools::Sim4::Exon objects. Call |
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this method repeatedly until an empty array is returned to get the |
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results for all alignments. |
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The $exon->seq_id() attribute will be set to the identifier of the |
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respective sequence for both sequences if A=4 was used in the sim4 |
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run, and otherwise for the second sequence only. If the output does |
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not contain the identifier, the filename stripped of path and |
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extension is used instead. In addition, the full filename |
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will be recorded for both features ($exon inherits off |
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Bio::SeqFeature::SimilarityPair) as tag 'filename'. The length |
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is accessible via the seqlength() attribute of $exon->query() and |
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$exon->est_hit(). |
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Note that this method is capable of dealing with outputs generated |
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with format 0,1,3, and 4 (via the A=n option to sim4). It |
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automatically determines which of the two sequences has been |
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reversed, and adjusts the coordinates for that sequence. It will |
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also detect whether the EST sequence(s) were given as first or as |
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second file to sim4, unless this has been specified at creation |
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time of the object. |
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Example : |
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Returns : An array of Bio::Tools::Sim4::Exon objects |
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Args : |
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=cut |
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sub parse_next_alignment { |
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my ($self) = @_; |
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my @exons = (); |
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my %seq1props = (); |
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my %seq2props = (); |
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# we refer to the properties of each seq by reference |
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my ($estseq, $genomseq, $to_reverse); |
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my $started = 0; |
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my $hit_direction = 1; |
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my $output_fmt = 3; # same as 0 and 1 (we cannot deal with A=2 produced |
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# output yet) |
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|
|
|
220
|
3
|
|
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|
|
12
|
while(defined($_ = $self->_readline())) { |
221
|
|
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|
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|
|
#chomp(); |
222
|
|
|
|
|
|
|
# |
223
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|
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|
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|
|
# bascially, each sim4 'hit' starts with seq1... |
224
|
|
|
|
|
|
|
# |
225
|
32
|
100
|
|
|
|
76
|
/^seq1/ && do { |
226
|
4
|
100
|
|
|
|
10
|
if($started) { |
227
|
1
|
|
|
|
|
14
|
$self->_pushback($_); |
228
|
1
|
|
|
|
|
2
|
last; |
229
|
|
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|
|
|
} |
230
|
3
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|
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|
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4
|
$started = 1; |
231
|
|
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|
|
|
|
|
232
|
|
|
|
|
|
|
# filename and length of seq 1 |
233
|
3
|
50
|
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|
|
16
|
/^seq1\s+=\s+(\S+)\,\s+(\d+)/ || |
234
|
|
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|
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|
|
$self->throw("Sim4 parsing error on seq1 [$_] line. Sorry!"); |
235
|
3
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|
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|
|
10
|
$seq1props{'filename'} = $1; |
236
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3
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|
|
8
|
$seq1props{'length'} = $2; |
237
|
3
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|
|
9
|
next; |
238
|
|
|
|
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|
|
}; |
239
|
28
|
100
|
|
|
|
47
|
/^seq2/ && do { |
240
|
|
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|
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|
|
# the second hit has also the database name in the >name syntax |
241
|
|
|
|
|
|
|
# (in brackets). |
242
|
3
|
50
|
|
|
|
16
|
/^seq2\s+=\s+(\S+)\s+\(>?(\S+)\s*\)\,\s+(\d+)/|| |
243
|
|
|
|
|
|
|
$self->throw("Sim4 parsing error on seq2 [$_] line. Sorry!"); |
244
|
3
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|
|
|
|
8
|
$seq2props{'filename'} = $1; |
245
|
3
|
|
|
|
|
7
|
$seq2props{'seqname'} = $2; |
246
|
3
|
|
|
|
|
7
|
$seq2props{'length'} = $3; |
247
|
3
|
|
|
|
|
6
|
next; |
248
|
|
|
|
|
|
|
}; |
249
|
25
|
50
|
|
|
|
45
|
if(/^>(\S+)\s*(.*)$/) { |
250
|
|
|
|
|
|
|
# output option was A=4, which not only gives the complete |
251
|
|
|
|
|
|
|
# description lines, but also causes the longer sequence to be |
252
|
|
|
|
|
|
|
# reversed if the second file contained one (genomic) sequence |
253
|
0
|
|
|
|
|
0
|
$seq1props{'seqname'} = $1; |
254
|
0
|
0
|
|
|
|
0
|
$seq1props{'description'} = $2 if $2; |
255
|
0
|
|
|
|
|
0
|
$output_fmt = 4; |
256
|
|
|
|
|
|
|
# we handle seq1 and seq2 both here |
257
|
0
|
0
|
0
|
|
|
0
|
if(defined($_ = $self->_readline()) && (/^>(\S+)\s*(.*)$/)) { |
258
|
0
|
|
|
|
|
0
|
$seq2props{'seqname'} = $1; # redundant, since already set above |
259
|
0
|
0
|
|
|
|
0
|
$seq2props{'description'} = $2 if $2; |
260
|
|
|
|
|
|
|
} |
261
|
0
|
|
|
|
|
0
|
next; |
262
|
|
|
|
|
|
|
} |
263
|
25
|
100
|
|
|
|
39
|
/^\(complement\)/ && do { |
264
|
1
|
|
|
|
|
2
|
$hit_direction = -1; |
265
|
1
|
|
|
|
|
2
|
next; |
266
|
|
|
|
|
|
|
}; |
267
|
|
|
|
|
|
|
# this matches |
268
|
|
|
|
|
|
|
# start-end (start-end) pctid% |
269
|
24
|
100
|
|
|
|
104
|
if(/(\d+)-(\d+)\s+\((\d+)-(\d+)\)\s+(\d+)%/) { |
270
|
18
|
|
|
|
|
47
|
$seq1props{'start'} = $1; |
271
|
18
|
|
|
|
|
34
|
$seq1props{'end'} = $2; |
272
|
18
|
|
|
|
|
36
|
$seq2props{'start'} = $3; |
273
|
18
|
|
|
|
|
30
|
$seq2props{'end'} = $4; |
274
|
18
|
|
|
|
|
28
|
my $pctid = $5; |
275
|
|
|
|
|
|
|
|
276
|
18
|
100
|
|
|
|
32
|
if(! defined($estseq)) { |
277
|
|
|
|
|
|
|
# for the first time here: need to set the references referring |
278
|
|
|
|
|
|
|
# to seq1 and seq2 |
279
|
3
|
100
|
|
|
|
10
|
if(! exists($self->{'_est_is_first'})) { |
280
|
|
|
|
|
|
|
# detect which one is the EST by looking at the lengths, |
281
|
|
|
|
|
|
|
# and assume that this holds throughout the entire result |
282
|
|
|
|
|
|
|
# file (i.e., when this method is called for the next |
283
|
|
|
|
|
|
|
# alignment, this will not be checked again) |
284
|
2
|
50
|
|
|
|
10
|
if($seq1props{'length'} > $seq2props{'length'}) { |
285
|
2
|
|
|
|
|
6
|
$self->{'_est_is_first'} = 0; |
286
|
|
|
|
|
|
|
} else { |
287
|
0
|
|
|
|
|
0
|
$self->{'_est_is_first'} = 1; |
288
|
|
|
|
|
|
|
} |
289
|
|
|
|
|
|
|
} |
290
|
3
|
50
|
|
|
|
7
|
if($self->{'_est_is_first'}) { |
291
|
0
|
|
|
|
|
0
|
$estseq = \%seq1props; |
292
|
0
|
|
|
|
|
0
|
$genomseq = \%seq2props; |
293
|
|
|
|
|
|
|
# if the EST is given first, A=4 selects the genomic |
294
|
|
|
|
|
|
|
# seq for being reversed (reversing the EST is default) |
295
|
0
|
0
|
|
|
|
0
|
$to_reverse = ($output_fmt == 4) ? $genomseq : $estseq; |
296
|
|
|
|
|
|
|
} else { |
297
|
3
|
|
|
|
|
6
|
$estseq = \%seq2props; |
298
|
3
|
|
|
|
|
8
|
$genomseq = \%seq1props; |
299
|
|
|
|
|
|
|
# if the EST is the second, A=4 does not change the |
300
|
|
|
|
|
|
|
# seq being reversed (always the EST is reversed) |
301
|
3
|
|
|
|
|
5
|
$to_reverse = $estseq; |
302
|
|
|
|
|
|
|
} |
303
|
|
|
|
|
|
|
} |
304
|
18
|
100
|
|
|
|
37
|
if($hit_direction == -1) { |
305
|
|
|
|
|
|
|
# we have to reverse the coordinates of one of both seqs |
306
|
10
|
|
|
|
|
14
|
my $tmp = $to_reverse->{'start'}; |
307
|
|
|
|
|
|
|
$to_reverse->{'start'} = |
308
|
10
|
|
|
|
|
25
|
$to_reverse->{'length'} - $to_reverse->{'end'} + 1; |
309
|
10
|
|
|
|
|
18
|
$to_reverse->{'end'} = $to_reverse->{'length'} - $tmp + 1; |
310
|
|
|
|
|
|
|
} |
311
|
|
|
|
|
|
|
# create and initialize the exon object |
312
|
|
|
|
|
|
|
my $exon = Bio::Tools::Sim4::Exon->new( |
313
|
|
|
|
|
|
|
'-start' => $genomseq->{'start'}, |
314
|
18
|
|
|
|
|
60
|
'-end' => $genomseq->{'end'}, |
315
|
|
|
|
|
|
|
'-strand' => $hit_direction); |
316
|
18
|
50
|
|
|
|
40
|
if(exists($genomseq->{'seqname'})) { |
317
|
0
|
|
|
|
|
0
|
$exon->seq_id($genomseq->{'seqname'}); |
318
|
|
|
|
|
|
|
} else { |
319
|
|
|
|
|
|
|
# take filename stripped of path as fall back |
320
|
18
|
|
|
|
|
469
|
my ($basename) = &File::Basename::fileparse($genomseq->{'filename'}, '\..*'); |
321
|
18
|
|
|
|
|
59
|
$exon->seq_id($basename); |
322
|
|
|
|
|
|
|
} |
323
|
|
|
|
|
|
|
$exon->feature1()->add_tag_value('filename', |
324
|
18
|
|
|
|
|
34
|
$genomseq->{'filename'}); |
325
|
|
|
|
|
|
|
# feature1 is supposed to be initialized to a Similarity object, |
326
|
|
|
|
|
|
|
# but we provide a safety net |
327
|
18
|
50
|
|
|
|
29
|
if($exon->feature1()->can('seqlength')) { |
328
|
18
|
|
|
|
|
28
|
$exon->feature1()->seqlength($genomseq->{'length'}); |
329
|
|
|
|
|
|
|
} else { |
330
|
|
|
|
|
|
|
$exon->feature1()->add_tag_value('SeqLength', |
331
|
0
|
|
|
|
|
0
|
$genomseq->{'length'}); |
332
|
|
|
|
|
|
|
} |
333
|
|
|
|
|
|
|
# create and initialize the feature wrapping the 'hit' (the EST) |
334
|
|
|
|
|
|
|
my $fea2 = Bio::SeqFeature::Similarity->new( |
335
|
|
|
|
|
|
|
'-start' => $estseq->{'start'}, |
336
|
18
|
|
|
|
|
51
|
'-end' => $estseq->{'end'}, |
337
|
|
|
|
|
|
|
'-strand' => 0, |
338
|
|
|
|
|
|
|
'-primary' => "aligning_EST"); |
339
|
18
|
50
|
|
|
|
32
|
if(exists($estseq->{'seqname'})) { |
340
|
18
|
|
|
|
|
37
|
$fea2->seq_id($estseq->{'seqname'}); |
341
|
|
|
|
|
|
|
} else { |
342
|
|
|
|
|
|
|
# take filename stripped of path as fall back |
343
|
|
|
|
|
|
|
my ($basename) = |
344
|
0
|
|
|
|
|
0
|
&File::Basename::fileparse($estseq->{'filename'}, '\..*'); |
345
|
0
|
|
|
|
|
0
|
$fea2->seq_id($basename); |
346
|
|
|
|
|
|
|
} |
347
|
18
|
|
|
|
|
42
|
$fea2->add_tag_value('filename', $estseq->{'filename'}); |
348
|
18
|
|
|
|
|
38
|
$fea2->seqlength($estseq->{'length'}); |
349
|
|
|
|
|
|
|
# store |
350
|
18
|
|
|
|
|
54
|
$exon->est_hit($fea2); |
351
|
|
|
|
|
|
|
# general properties |
352
|
18
|
|
|
|
|
32
|
$exon->source_tag($self->analysis_method()); |
353
|
18
|
|
|
|
|
38
|
$exon->percentage_id($pctid); |
354
|
18
|
|
|
|
|
30
|
$exon->score($exon->percentage_id()); |
355
|
|
|
|
|
|
|
# push onto array |
356
|
18
|
|
|
|
|
28
|
push(@exons, $exon); |
357
|
18
|
|
|
|
|
51
|
next; # back to while loop |
358
|
|
|
|
|
|
|
} |
359
|
|
|
|
|
|
|
} |
360
|
3
|
|
|
|
|
18
|
return @exons; |
361
|
|
|
|
|
|
|
} |
362
|
|
|
|
|
|
|
|
363
|
|
|
|
|
|
|
=head2 next_exonset |
364
|
|
|
|
|
|
|
|
365
|
|
|
|
|
|
|
Title : next_exonset |
366
|
|
|
|
|
|
|
Usage : $exonset = $sim4_result->parse_next_exonset; |
367
|
|
|
|
|
|
|
print "Exons start at ", $exonset->start(), |
368
|
|
|
|
|
|
|
"and end at ", $exonset->end(), "\n"; |
369
|
|
|
|
|
|
|
foreach $exon ($exonset->sub_SeqFeature()) { |
370
|
|
|
|
|
|
|
# do something |
371
|
|
|
|
|
|
|
} |
372
|
|
|
|
|
|
|
Function: Parses the next alignment of the Sim4 result file and returns the |
373
|
|
|
|
|
|
|
set of exons as a container of features. The container is itself |
374
|
|
|
|
|
|
|
a Bio::SeqFeature::Generic object, with the Bio::Tools::Sim4::Exon |
375
|
|
|
|
|
|
|
objects as sub features. Start, end, and strand of the container |
376
|
|
|
|
|
|
|
will represent the total region covered by the exons of this set. |
377
|
|
|
|
|
|
|
|
378
|
|
|
|
|
|
|
See the documentation of parse_next_alignment() for further |
379
|
|
|
|
|
|
|
reference about parsing and how the information is stored. |
380
|
|
|
|
|
|
|
|
381
|
|
|
|
|
|
|
Example : |
382
|
|
|
|
|
|
|
Returns : An Bio::SeqFeature::Generic object holding Bio::Tools::Sim4::Exon |
383
|
|
|
|
|
|
|
objects as sub features. |
384
|
|
|
|
|
|
|
Args : |
385
|
|
|
|
|
|
|
|
386
|
|
|
|
|
|
|
=cut |
387
|
|
|
|
|
|
|
|
388
|
|
|
|
|
|
|
sub next_exonset { |
389
|
3
|
|
|
3
|
1
|
781
|
my $self = shift; |
390
|
3
|
|
|
|
|
5
|
my $exonset; |
391
|
|
|
|
|
|
|
|
392
|
|
|
|
|
|
|
# get the next array of exons |
393
|
3
|
|
|
|
|
10
|
my @exons = $self->parse_next_alignment(); |
394
|
3
|
50
|
|
|
|
12
|
unless( @exons ) { |
395
|
0
|
0
|
|
|
|
0
|
return if eof($self->_fh); |
396
|
0
|
|
|
|
|
0
|
return $self->next_exonset; |
397
|
|
|
|
|
|
|
} |
398
|
|
|
|
|
|
|
# create the container of exons as a feature object itself, with the |
399
|
|
|
|
|
|
|
# data of the first exon for initialization |
400
|
3
|
|
|
|
|
10
|
$exonset = Bio::SeqFeature::Generic->new('-start' => $exons[0]->start(), |
401
|
|
|
|
|
|
|
'-end' => $exons[0]->end(), |
402
|
|
|
|
|
|
|
'-strand' => $exons[0]->strand(), |
403
|
|
|
|
|
|
|
'-primary' => "ExonSet"); |
404
|
3
|
|
|
|
|
11
|
$exonset->source_tag($exons[0]->source_tag()); |
405
|
3
|
|
|
|
|
9
|
$exonset->seq_id($exons[0]->seq_id()); |
406
|
|
|
|
|
|
|
# now add all exons as sub features, with enabling EXPANsion of the region |
407
|
|
|
|
|
|
|
# covered in total |
408
|
3
|
|
|
|
|
9
|
foreach my $exon (@exons) { |
409
|
18
|
|
|
|
|
34
|
$exonset->add_sub_SeqFeature($exon, 'EXPAND'); |
410
|
|
|
|
|
|
|
} |
411
|
3
|
|
|
|
|
20
|
return $exonset; |
412
|
|
|
|
|
|
|
} |
413
|
|
|
|
|
|
|
|
414
|
|
|
|
|
|
|
=head2 next_feature |
415
|
|
|
|
|
|
|
|
416
|
|
|
|
|
|
|
Title : next_feature |
417
|
|
|
|
|
|
|
Usage : while($exonset = $sim4->next_feature()) { |
418
|
|
|
|
|
|
|
# do something |
419
|
|
|
|
|
|
|
} |
420
|
|
|
|
|
|
|
Function: Does the same as L. See there for documentation of |
421
|
|
|
|
|
|
|
the functionality. Call this method repeatedly until FALSE is |
422
|
|
|
|
|
|
|
returned. |
423
|
|
|
|
|
|
|
|
424
|
|
|
|
|
|
|
The returned object is actually a SeqFeatureI implementing object. |
425
|
|
|
|
|
|
|
This method is required for classes implementing the |
426
|
|
|
|
|
|
|
SeqAnalysisParserI interface, and is merely an alias for |
427
|
|
|
|
|
|
|
next_exonset() at present. |
428
|
|
|
|
|
|
|
|
429
|
|
|
|
|
|
|
Example : |
430
|
|
|
|
|
|
|
Returns : A Bio::SeqFeature::Generic object. |
431
|
|
|
|
|
|
|
Args : |
432
|
|
|
|
|
|
|
|
433
|
|
|
|
|
|
|
=cut |
434
|
|
|
|
|
|
|
|
435
|
|
|
|
|
|
|
sub next_feature { |
436
|
0
|
|
|
0
|
1
|
|
my ($self,@args) = @_; |
437
|
|
|
|
|
|
|
# even though next_exonset doesn't expect any args (and this method |
438
|
|
|
|
|
|
|
# does neither), we pass on args in order to be prepared if this changes |
439
|
|
|
|
|
|
|
# ever |
440
|
0
|
|
|
|
|
|
return $self->next_exonset(@args); |
441
|
|
|
|
|
|
|
} |
442
|
|
|
|
|
|
|
|
443
|
|
|
|
|
|
|
1; |