# $Id: ace.pm 16123 2009-09-17 12:57:27Z cjfields $ # ## BioPerl module for Bio::Assembly::IO::ace # # Copyright by Robson F. de Souza # # You may distribute this module under the same terms as perl itself # POD documentation - main docs before the code =head1 NAME Bio::Assembly::IO::ace - module to load ACE files from various assembly programs =head1 SYNOPSIS # Building an input stream use Bio::Assembly::IO; # Assembly loading methods $io = Bio::Assembly::IO->new( -file=>"SGC0-424.fasta.screen.ace.1", -format=>"ace" ); $assembly = $io->next_assembly; =head1 DESCRIPTION This package loads the standard ACE files generated by various assembly programs (Phrap, CAP3, Newbler, Arachne, ...). It was written to be used as a driver module for Bio::Assembly::IO input/output. =head2 Implemention Assemblies are loaded into Bio::Assembly::Scaffold objects composed by Bio::Assembly::Contig and Bio::Assembly::Singlet objects. Only the ACE file is used, so if you need singlets, make sure that they are present in the ACE file. A description of the ACE format is available at http://www.cbcb.umd.edu/research/contig_representation.shtml#ACE In addition to default "_aligned_coord:$seqID" feature class from Bio::Assembly::Contig, contig objects loaded by this module will have the following special feature classes in their feature collection: "_align_clipping:$seqID" : location of subsequence in sequence $seqID which is aligned to the contig. If no feature containing this tag is present the read is considered low quality by consed "_quality_clipping:$seqID" : location of good quality subsequence for sequence $seqID "consensus tags", as they are called in Consed's documentation, is equivalent to a bioperl sequence feature and, therefore, are added to the feature collection using their type field (see Consed's README.txt file) as primary tag. "read tags" are also sequence features and are stored as sub_SeqFeatures of the sequence's coordinate feature (the corresponding "_aligned_coord:$seqID" feature, easily accessed through get_seq_coord() method). "whole assembly tags" have no start and end, as they are not associated to any particular sequence in the assembly, and are added to the assembly's annotation collection using "whole assembly" as tag. =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 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 - Robson Francisco de Souza Email rfsouza@citri.iq.usp.br =head1 APPENDIX The rest of the documentation details each of the object methods. Internal methods are usually preceded with a _ =cut package Bio::Assembly::IO::ace; use strict; use Bio::Assembly::Scaffold; use Bio::Assembly::Contig; use Bio::Assembly::Singlet; use Bio::LocatableSeq; use Bio::Annotation::SimpleValue; use Bio::Seq::Quality; use Bio::SeqIO; use Bio::SeqFeature::Generic; use base qw(Bio::Assembly::IO); =head1 Parser methods =head2 next_assembly Title : next_assembly Usage : $unigene = $stream->next_assembly() Function: returns the next assembly in the stream Returns : Bio::Assembly::Scaffold object Args : NONE =cut sub next_assembly { my $self = shift; # Object reference my $lingering_read; local $/ = "\n"; # Resetting assembly data structure my $assembly = Bio::Assembly::Scaffold->new(); # Looping over all ACE file lines my ($contigOBJ,$read_name); my $read_data = {}; # Temporary holder for read data while ($_ = $self->_readline) { # By now, ACE files hold a single assembly chomp; # Loading assembly information (ASsembly field) #(/^AS\s+(\d+)\s+(\d+)/) && do { # $assembly->_set_nof_contigs($1); # $assembly->_set_nof_sequences_in_contigs($2); #}; # Loading contig sequence (COntig sequence field) (/^CO\s(\w+)\s(\d+)\s(\d+)\s(\d+)\s(\w+)/xms) && do { # New contig found! my $contigID = $1; # Contig ID #my $nof_bases = $2; # Contig length in base pairs my $nof_reads = $3; # Number of reads in this contig #my $nof_segments = $4; # Number of read segments selected for consensus assembly my $ori = $5; # 'C' if contig was complemented or U if not (default) $ori = ($ori eq 'U' ? 1 : -1); # Create a singlet if necessary if ($nof_reads < 1) { $self->throw("Expecting a strictly positive number of reads in a contig"); } elsif ($nof_reads == 1) { # This is a singlet $contigOBJ = Bio::Assembly::Singlet->new( ); } else { # $nof_reads > 1 # This is a contig $contigOBJ = Bio::Assembly::Contig->new( -id => $contigID ); $contigOBJ->strand($ori); } my $consensus_sequence = undef; while ($_ = $self->_readline) { # Looping over contig lines chomp; # Drop (\n) on current line last if (/^$/); # Stop if empty line (contig end) is found s/\*/-/g; # Forcing '-' as gap symbol $consensus_sequence .= $_; } $consensus_sequence = Bio::LocatableSeq->new( -seq => $consensus_sequence, -start => 1, -id => $contigID ); if ($contigOBJ->isa('Bio::Assembly::Singlet')) { # a singlet $contigOBJ->seqref($consensus_sequence); $assembly->add_singlet($contigOBJ); } else { # a contig $contigOBJ->set_consensus_sequence($consensus_sequence); $assembly->add_contig($contigOBJ); } }; # Loading contig qualities... (Base Quality field) /^BQ/ && do { my $consensus = $contigOBJ->get_consensus_sequence()->seq(); my ($i,$j,@tmp); my @quality = (); $j = 0; while ($_ = $self->_readline) { chomp; last if (/^$/); @tmp = grep { /^\d+$/ } split(/\s+/); $i = 0; my $previous = 0; my $next = 0; while ($i<=$#tmp) { # IF base is a gap, quality is the average for neighbouring sites if (substr($consensus,$j,1) eq '-') { $previous = $tmp[$i-1] unless ($i == 0); if ($i < $#tmp) { $next = $tmp[$i+1]; } else { $next = 0; } push(@quality,int(($previous+$next)/2)); } else { push(@quality,$tmp[$i]); $i++; } $j++; } } my $qual = Bio::Seq::Quality->new( -qual=>join(" ",@quality), -id=>$contigOBJ->id() ); $contigOBJ->set_consensus_quality($qual); }; # Loading read info... (Assembled From field) /^AF (\S+) (C|U) (-*\d+)/ && do { $read_name = $1; my $ori = $2; $read_data->{$read_name}{'padded_start'} = $3; # aligned start $ori = ( $ori eq 'U' ? 1 : -1); $read_data->{$read_name}{'strand'} = $ori; }; # Loading base segments definitions (Base Segment field) # They indicate which read segments were used to calculate the consensus # Safe to ignore # /^BS (\d+) (\d+) (\S+)/ && do { # if (exists($self->{'contigs'}[$contig]{'reads'}{$3}{'segments'})) { # $self->{'contigs'}[$contig]{'reads'}{$3}{'segments'} .= " " . $1 . " " . $2; # } else { # $self->{'contigs'}[$contig]{'reads'}{$3}{'segments'} = $1 . " " . $2 # } # }; # Loading reads... (ReaD sequence field) /^RD (\S+) (-*\d+) (\d+) (\d+)/ && do { next if $contigOBJ->isa('Bio::Assembly::Singlet'); $read_name = $1; $read_data->{$read_name}{'length'} = $2; # number_of_padded_bases $read_data->{$read_name}{'contig'} = $contigOBJ; # $read_data->{$read_name}{'number_of_read_info_items'} = $3; # $read_data->{$read_name}{'number_of_tags'} = $4; my $read_sequence; while ($_ = $self->_readline) { chomp; last if (/^$/); s/\*/-/g; # Forcing '-' as gap symbol $read_sequence .= $_; # aligned read sequence } my $read = Bio::LocatableSeq->new( -seq=>$read_sequence, -start=>1, -strand=>$read_data->{$read_name}{'strand'}, -id=>$read_name, -primary_id=>$read_name, -alphabet=>'dna' ); $lingering_read = $read; # Adding read location and sequence to contig ("gapped consensus" coordinates) my $padded_start = $read_data->{$read_name}{'padded_start'}; my $padded_end = $padded_start + $read_data->{$read_name}{'length'} - 1; my $coord = Bio::SeqFeature::Generic->new( -start=>$padded_start, -end=>$padded_end, -strand=>$read_data->{$read_name}{'strand'}, -tag => { 'contig' => $contigOBJ->id } ); $contigOBJ->set_seq_coord($coord,$read); }; # Loading read trimming and alignment ranges... /^QA (-?\d+) (-?\d+) (-?\d+) (-?\d+)/ && do { next if $contigOBJ->isa('Bio::Assembly::Singlet'); my ($qual_start, $qual_end, $align_start, $align_end) = ($1, $2, $3, $4); unless ($align_start == -1 && $align_end == -1) { $align_start = $contigOBJ->change_coord("aligned $read_name",'gapped consensus',$align_start); $align_end = $contigOBJ->change_coord("aligned $read_name",'gapped consensus',$align_end); my $align_feat = Bio::SeqFeature::Generic->new( -start=>$align_start, -end=>$align_end, -strand=>$read_data->{$read_name}{'strand'}, -primary=>"_align_clipping:$read_name"); $align_feat->attach_seq( $contigOBJ->get_seq_by_name($read_name) ); $contigOBJ->add_features([ $align_feat ], 0); } unless ($qual_start == -1 && $qual_end == -1) { $qual_start = $contigOBJ->change_coord("aligned $read_name",'gapped consensus',$qual_start); $qual_end = $contigOBJ->change_coord("aligned $read_name",'gapped consensus',$qual_end); my $qual_feat = Bio::SeqFeature::Generic->new( -start=>$qual_start, -end=>$qual_end, -strand=>$read_data->{$read_name}{'strand'}, -primary=>"_quality_clipping:$read_name" ); $qual_feat->attach_seq( $contigOBJ->get_seq_by_name($read_name) ); $contigOBJ->add_features([ $qual_feat ], 0); } }; # Loading read description (DeScription fields) # chad was here! easter 2004. # lingering read is a locatableseq. is there a better way to do this? # i am simply adding more keys to the locatableseq # Should that no be put in a Bio::SeqFeature::Generic object? #/^DS / && do { # /CHEM: (\S+)/ && do { # $lingering_read->{'chemistry'} = $1; # }; # /CHROMAT_FILE: (\S+)/ && do { # $lingering_read->{'chromatfilename'} = $1; # }; # /DIRECTION: (\w+)/ && do { # my $ori = $1; # if ($ori eq 'rev') { $ori = 'C' } # elsif ($ori eq 'fwd') { $ori = 'U' } # $lingering_read->{'strand'} = $ori; # }; # /DYE: (\S+)/ && do { # $lingering_read->{'dye'} = $1; # }; # /PHD_FILE: (\S+)/ && do { # $lingering_read->{'phdfilename'} = $1; # }; # /TEMPLATE: (\S+)/ && do { # $lingering_read->{'template'} = $1; # }; # /TIME: (\S+ \S+ \d+ \d+\:\d+\:\d+ \d+)/ && do { # $lingering_read->{'phd_time'} = $1; # }; #}; # Loading Contig Tags (a.k.a. Bioperl features) /^CT\s*\{/ && do { my ($contigID,$type,$source,$start,$end,$date) = split(' ',$self->_readline); my %tags = (source => $source, creation_date => $date); my $tag_type = 'extra_info'; while ($_ = $self->_readline) { if (/COMMENT\s*\{/) { $tag_type = 'comment'; } elsif (/C\}/) { $tag_type = 'extra_info'; } elsif (/\}/) { last; } else { $tags{$tag_type} .= "$_"; } } my $contig_tag = Bio::SeqFeature::Generic->new( -start=>$start, -end=>$end, -primary=>$type, -tag=>\%tags ); my $contig = $assembly->get_contig_by_id($contigID); $self->throw("Cannot add feature to unknown contig '$contigID'") unless defined $contig; $contig->add_features([ $contig_tag ],1); }; # Loading Read Tags /^RT\s*\{/ && do { next if $contigOBJ->isa('Bio::Assembly::Singlet'); my ($readID,$type,$source,$start,$end,$date) = split(' ',$self->_readline); my $extra_info = undef; while ($_ = $self->_readline) { last if (/\}/); $extra_info .= $_; } $start = $contigOBJ->change_coord("aligned $read_name",'gapped consensus',$start); $end = $contigOBJ->change_coord("aligned $read_name",'gapped consensus',$end); my $read_tag = Bio::SeqFeature::Generic->new( -start=>$start, -end=>$end, -primary=>$type, -tag=>{ 'source' => $source, 'creation_date' => $date, 'extra_info' => $extra_info } ); my $contig = $read_data->{$readID}{'contig'}; my $coord = $contig->get_seq_coord( $contig->get_seq_by_name($readID) ); $coord->add_sub_SeqFeature($read_tag); }; # Loading Whole Assembly tags /^WA\s*\{/ && do { my ($type,$source,$date) = split(' ',$self->_readline); my $extra_info = undef; while ($_ = $self->_readline) { last if (/\}/); $extra_info = $_; } #? push(@line,\@extra_info); my $assembly_tag = join(" ","TYPE: ",$type,"PROGRAM:",$source, "DATE:",$date,"DATA:",$extra_info); $assembly_tag = Bio::Annotation::SimpleValue->new(-value=>$assembly_tag); $assembly->annotation->add_Annotation('whole assembly',$assembly_tag); }; } # while ($_ = $self->_readline) # hmm. what about singlets? # Singlets are expected to be present in the ACE file, not elsewhere #my $singletsfilename = $self->file(); #$singletsfilename =~ s/\.ace.*$/.singlets/; #$singletsfilename =~ s/\new( -file => "<$singletsfilename", # -format => 'fasta' ); # my $adder; # while (my $seq = $singlets_fh->next_seq()) { # # find the name of this singlet and attempt to get the phd from phd_dir instead # my ($phdfilename,$chromatfilename) = qw(unset unset); # if ($seq->desc() =~ /PHD_FILE: (\S+)/) { # $phdfilename = $1; # } # if ($seq->desc() =~ /CHROMAT_FILE: (\S+)/) { # $chromatfilename = $1; # } # # should chromatfile and phdfile be put in a Bio::SeqFeature::Generic object? # (my $phdfile = $singletsfilename) =~ s/edit_dir.*//; # $phdfile .= "phd_dir/$phdfilename"; # my $singlet = Bio::Assembly::Singlet->new(); # if (-f $phdfile) { # # print STDERR ("Reading singlet data from this phdfile ($phdfile)\n"); # my $phd_fh = Bio::SeqIO->new( -file => "<$phdfile", -format => 'phd'); # my $swq = $phd_fh->next_seq(); # $adder = $swq; # } else { # $adder = $seq; # } # $singlet->seqref($adder); # $assembly->add_singlet($singlet); # } #} $assembly->update_seq_list(); $assembly->get_nof_contigs ? return $assembly : return; } =head2 write_assembly Title : write_assembly Usage : $ass_io->write_assembly($assembly) Function: Write the assembly object in ACE compatible format Returns : 1 on success, 0 for error Args : A Bio::Assembly::Scaffold object =cut sub write_assembly { my $self = shift; $self->throw_not_implemented(); } 1; __END__