# $Id: Results.pm 16123 2009-09-17 12:57:27Z cjfields $ # # BioPerl module for Bio::Tools::Spidey::Results # # Please direct questions and support issues to # # Cared for by Ryan Golhar # # You may distribute this module under the same terms as perl itself # POD documentation - main docs before the code =head1 NAME Bio::Tools::Spidey::Results - Results of a Spidey run =head1 SYNOPSIS use Bio::Tools::Spidey::Results; my $spidey = Bio::Tools::Spidey::Results->new(-file => 'result.spidey' ); # or my $spidey = Bio::Tools::Spidey::Results->new( -fh => \*INPUT ); # get the exons before doing anything else my $exonset = $spidey->next_exonset(); # parse the results my @exons = $exonset->sub_SeqFeature(); print "Total no of Exons: ", scalar(@exons), "\n"; print "Genomic sequence length: ", $spidey->genomic_dna_length(), "\n"; # $exonset is-a Bio::SeqFeature::Generic with Bio::Tools::Spidey::Exons # as sub features print "Delimited on sequence ", $exonset->seq_id(), " from ", $exonset->start(), " to ", $exonset->end(), "\n"; foreach my $exon ( $exonset->sub_SeqFeature() ) { # $exon is-a Bio::SeqFeature::FeaturePair print "Exon from ", $exon->start, " to ", $exon->end, " on strand ", $exon->strand(), "\n"; # you can get out what it matched using the est_hit attribute my $homol = $exon->est_hit(); print "Matched to sequence ", $homol->seq_id, " at ", $homol->start," to ", $homol->end, "\n"; } # essential if you gave a filename at initialization (otherwise # the file stays open) $spidey->close(); =head1 DESCRIPTION The spidey module provides a parser and results object for spidey output. The spidey results are specialised types of SeqFeatures, meaning you can add them to AnnSeq objects fine, and manipulate them in the "normal" seqfeature manner. The spidey Exon objects are Bio::SeqFeature::FeaturePair inherited objects. The $esthit = $exon-Eest_hit() is the alignment as a feature on the matching object (normally, a cDNA), in which the start/end points are where the hit lies. To make this module work sensibly you need to run spidey -i genomic.fasta -m cDNA.fasta =head1 FEEDBACK =head2 Mailing Lists User feedback is an integral part of the evolution of this and other Bioperl modules. Send your comments and suggestions preferably to one of the Bioperl mailing lists. Your participation is much appreciated. bioperl-l@bioperl.org - General discussion http://bioperl.org/wiki/Mailing_lists - About the mailing lists =head2 Support Please direct usage questions or support issues to the mailing list: I rather than to the module maintainer directly. Many experienced and reponsive experts will be able look at the problem and quickly address it. Please include a thorough description of the problem with code and data examples if at all possible. =head2 Reporting Bugs Report bugs to the Bioperl bug tracking system to help us keep track the bugs and their resolution. Bug reports can be submitted via the web: http://bugzilla.open-bio.org/ =head1 AUTHOR - Ryan Golhar Email golharam@umdnj.edu =head1 APPENDIX The rest of the documentation details each of the object methods. Internal methods are usually preceded with a _ =cut # Let the code begin... package Bio::Tools::Spidey::Results; use strict; use File::Basename; use Bio::Root::Root; use Bio::Tools::Spidey::Exon; use base qw(Bio::Tools::AnalysisResult); sub _initialize_state { my($self,@args) = @_; # call the inherited method first my $make = $self->SUPER::_initialize_state(@args); # my ($est_is_first) = $self->_rearrange([qw(ESTFIRST)], @args); # delete($self->{'_est_is_first'}); # $self->{'_est_is_first'} = $est_is_first if(defined($est_is_first)); $self->analysis_method("Spidey"); } =head2 analysis_method Usage : $spidey->analysis_method(); Purpose : Inherited method. Overridden to ensure that the name matches /Spidey/i. Returns : String Argument : n/a =cut #------------- sub analysis_method { #------------- my ($self, $method) = @_; if($method && ($method !~ /Spidey/i)) { $self->throw("method $method not supported in " . ref($self)); } return $self->SUPER::analysis_method($method); } =head2 parse_next_alignment Title : parse_next_alignment Usage : @exons = $spidey_result->parse_next_alignment; foreach $exon (@exons) { # do something } Function: Parses the next alignment of the Spidey result file and returns the found exons as an array of Bio::Tools::Spidey::Exon objects. Call this method repeatedly until an empty array is returned to get the results for all alignments. Example : Returns : An array of Bio::Tools::Spidey::Exon objects Args : =cut sub parse_next_alignment { my ($self) = @_; # for strand 1 = plus, -1 = minus my ($started,$version,$strand, $exoncount) = (0,0,0,-1); my (%seq1props,%seq2props,@exons); # we refer to the properties of each seq by reference while(defined($_ = $self->_readline())) { chomp; # # bascially, parse a Spidey result... # # matches: --SPIDEY version 1.40-- if( /^--SPIDEY\s+version\s+(\d+\.\d+)--/) { if($started) { $self->_pushback($_); return \@exons; } $version = $1; if ($version != 1.40) { $self->throw("Spidey parser only designed to work with Spidey v1.40\n"); } $started = 1; } elsif (/^Genomic:\s+(\S+)\s.*,\s+(\d+)\sbp$/ ) { # matches: Genomic: lcl|some_name other information, 1234 bp # $seq1props{'filename'} = $1; $seq1props{'seqname'} = $1; $seq1props{'length'} = $2; $self->genomic_dna_length($seq1props{'length'}); } elsif( /^mRNA:\s+(\S+)\s.*,(?:\s+mRNA\s+sequence,)?\s(\d+)\sbp$/ ) { # matches: mRNA: # $seq2props{'filename'} = $1; $seq2props{'seqname'} = $1; $seq2props{'length'} = $2; } elsif( /^Strand:/ ) { if (/plus/) { $strand = 1; } else { $strand = -1; } } elsif( /^Number of exons: (\d+)/ ) { $exoncount = $1; my ($genomic_start, $genomic_stop, $cdna_start, $cdna_stop, $id, $mismatches, $gaps, $splice_donor, $splice_acceptor, $uncertain); # the next $exoncount lines contains information # about the matches of each exon. we should parse # this information here for (my $ec = 1; $ec <= $exoncount; $ec++) { if (defined($_ = $self->_readline())) { chomp; if (/^Exon\s$ec[\(\)-]*:\s(\d+)-(\d+)\s\(gen\)\s+(\d+)-(\d+)\s\(mRNA\)\s+id\s([\d\.inf-]+)%\s+mismatches\s(\d+)\s+gaps\s(\d+)\s+splice\ssite\s\(d\s+a\):\s(\d+)\s+(\d+)\s*(\w*)/) { $genomic_start = $1; $genomic_stop = $2; $cdna_start = $3; $cdna_stop = $4; $id = $5; $mismatches = $6; $gaps = $7; $splice_donor = $8; $splice_acceptor = $9; $uncertain = $10; } else { $self->throw( "Failed to match anything:\n$_\n"); } my $exon = Bio::Tools::Spidey::Exon->new (-start => $genomic_start, -end => $genomic_stop, -strand => $strand); $exon->seq_id($seq1props{'seqname'}); # feature1 is supposed to be initialized to a Similarity object, but we provide a safety net if ($exon->feature1->can('seqlength')) { $exon->feature1->seqlength($seq1props{'length'}); } else { $exon->feature1->add_tag_value('seqlength', $seq1props{'length'}); } # create and initialize the feature wrapping the 'hit' (the cDNA) my $fea2 = Bio::SeqFeature::Similarity->new (-start => $cdna_start, -end => $cdna_stop, -strand => $strand, -seq_id => $seq2props{'seqname'}, -primary => "aligning_cDNA"); $fea2->seqlength($seq2props{'length'}); # store $exon->est_hit($fea2); # general properties $exon->source_tag($self->analysis_method()); $exon->percentage_id($5); $exon->mismatches($6); $exon->gaps($7); $exon->donor($8); $exon->acceptor($9); # push onto array push(@exons, $exon); } else { $self->throw("Unexpected end of file reached\n"); } } } elsif( /^Number of splice sites:\s+(\d+)/ ) { $self->splicesites($1); } elsif( /^mRNA coverage:\s+(\d+)%/ ) { $self->est_coverage($1); } elsif(/^overall percent identity:\s+([\d\.]+)%/ ) { $self->overall_percentage_id($1); } elsif(/^Missing mRNA ends:\s+(\w+)/ ) { $self->missing_mrna_ends($1); } elsif( /^Exon (\d+): (\d+)-(\d+) \(gen\)\s+(\d+)-(\d+) \(mRNA\)/ ) { my ($exon_num, $gen_start, $gen_stop, $cdna_start, $cdna_stop); $exon_num = $1; $gen_start = $2; $gen_stop = $3; $cdna_start = $4; $cdna_stop = $5; } elsif( /No alignment found/ ) { return []; } else { #$self->debug("unmatched $_\n"); } } # Typical format: # Exon 1: 36375798-36375691 (gen) 1-108 (mRNA) # # # CCTCTTTTTCTTTGCAGGGTATATACCCAGTTACTTAGACAAGGATGAGCTATGTGTAGT # | |||||||||||||||||||||||||||||||||||||||||||||| # ATGTCAGGGTATATACCCAGTTACTTAGACAAGGATGAGCTATGTGTAGT # M S G Y I P S Y L D K D E L C V V # # # ATGTGGGGACAAAGCCACCGGATATCATTATCGCTGCATCACTTGTGAAGGTTGCAAGGT # |||||||||||||||||||||||||||||||||||||||||||||||||||||||||| # ATGTGGGGACAAAGCCACCGGATATCATTATCGCTGCATCACTTGTGAAGGTTGCAAG # C G D K A T G Y H Y R C I T C E G C K # # # AAATGGCA # @exons ? return \@exons : return ; } =head2 next_exonset Title : next_exonset Usage : $exonset = $spidey_result->parse_next_exonset; print "Exons start at ", $exonset->start(), "and end at ", $exonset->end(), "\n"; for $exon ($exonset->sub_SeqFeature()) { # do something } Function: Parses the next alignment of the Spidey result file and returns the set of exons as a container of features. The container is itself a Bio::SeqFeature::Generic object, with the Bio::Tools::Spidey::Exon objects as sub features. Start, end, and strand of the container will represent the total region covered by the exons of this set. See the documentation of parse_next_alignment() for further reference about parsing and how the information is stored. Example : Returns : An Bio::SeqFeature::Generic object holding Bio::Tools::Spidey::Exon objects as sub features. Args : =cut sub next_exonset { my $self = shift; my $exonset; # get the next array of exons my $exons = $self->parse_next_alignment(); if( ! defined $exons ) { $self->warn("No exons returned"); return; } if( @$exons == 0 ) { return Bio::SeqFeature::Generic->new(); } # create the container of exons as a feature object itself, with the # data of the first exon for initialization $exonset = Bio::SeqFeature::Generic->new('-start' => $exons->[0]->start(), '-end' => $exons->[-1]->end(), '-strand' => $exons->[0]->strand(), '-primary' => "ExonSet"); $exonset->source_tag($exons->[0]->source_tag()); $exonset->seq_id($exons->[0]->seq_id()); # now add all exons as sub features, with enabling EXPANsion of the region # covered in total foreach my $exon (@$exons) { $exonset->add_sub_SeqFeature($exon, 'EXPAND'); } return $exonset; } =head2 next_feature Title : next_feature Usage : while($exonset = $spidey->next_feature()) { # do something } Function: Does the same as L. See there for documentation of the functionality. Call this method repeatedly until FALSE is returned. The returned object is actually a SeqFeatureI implementing object. This method is required for classes implementing the SeqAnalysisParserI interface, and is merely an alias for next_exonset() at present. Example : Returns : A Bio::SeqFeature::Generic object. Args : =cut sub next_feature { my ($self,@args) = @_; # even though next_exonset doesn't expect any args (and this method # does neither), we pass on args in order to be prepared if this changes # ever return $self->next_exonset(@args); } =head2 genomic_dna_length Title : genomic_dna_length Usage : $spidey->genomic_dna_length(); Function: Returns the length of the genomic DNA used in this Spidey result Example : Returns : An integer value. Args : =cut sub genomic_dna_length { my ($self, @args) = @_; my $val; if(@args) { $val = shift(@args); $self->{'genomic_dna_length'} = $val; } else { $val = $self->{'genomic_dna_length'}; } return $val; } =head2 splicesites Title : splicesites Usage : $spidey->splicesites(); Function: Returns the number of splice sites found in this Spidey result Example : Returns : An integer value. Args : =cut sub splicesites { my ($self, @args) = @_; my $val; if(@args) { $val = shift(@args); $self->{'splicesites'} = $val; } else { $val = $self->{'splicesites'}; } return $val; } =head2 est_coverage Title : est_coverage Usage : $spidey->est_coverage(); Function: Returns the percent of est coverage in this Spidey result Example : Returns : An integer value. Args : =cut sub est_coverage { my ($self, @args) = @_; my $val; if(@args) { $val = shift(@args); $self->{'est_coverage'} = $val; } else { $val = $self->{'est_coverage'}; } return $val; } =head2 overall_percentage_id Title : overall_percentage_id Usage : $spidey->overall_percentage_id(); Function: Returns the overall percent id in this Spidey result Example : Returns : An float value. Args : =cut sub overall_percentage_id { my ($self, @args) = @_; my $val; if(@args) { $val = shift(@args); $self->{'overall_percentage_id'} = $val; } else { $val = $self->{'overall_percentage_id'}; } return $val; } =head2 missing_mrna_ends Title : missing_mrna_ends Usage : $spidey->missing_mrna_ends(); Function: Returns left/right/neither from Spidey Example : Returns : A string value. Args : =cut sub missing_mrna_ends { my ($self, @args) = @_; my $val; if(@args) { $val = shift(@args); $self->{'missing_mrna_ends'} = $val; } else { $val = $self->{'missing_mrna_ends'}; } return $val; } 1;