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author | cpt |
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date | Mon, 05 Jun 2023 02:47:24 +0000 |
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#!/usr/bin/env perl use strict; use warnings; use Storable; use Bio::SearchIO; use Bio::SeqIO; use Bio::Tools::CodonTable; use Data::Dumper; use File::Temp qw/tempfile tempdir/; use CPT::GalaxyGetOpt; my $ggo = CPT::GalaxyGetOpt->new(); my $options = $ggo->getOptions( 'options' => [ [ 'file', 'Input file', { validate => 'File/Input', multiple => 1, required => 1, file_format => ['genbank', 'txt'], } ], ], 'outputs' => [ [ 'cpt_psm_object', 'Output PSM Object', { validate => 'File/Output', required => 1, default => 'cpt_psm', data_format => 'text/plain', default_format => 'TXT' } ], ], 'defaults' => [ 'appid' => 'PSM.Prep', 'appname' => 'PSM Prep', 'appdesc' => 'prepares data for the PSM Plotter', 'appvers' => '1.94.2', ], 'tests' => [], ); use CPT::Bio; my $bio = CPT::Bio->new(); my @genbank_files = @{$options->{file}}; my %data = ( file_list => [], ); my $GLOBAL_UNLINK_VAR = 1; my $tempdir = tempdir('cpt.psm2.XXXXXXX',CLEANUP => $GLOBAL_UNLINK_VAR); use CPT::Util::CRC64; my $crc = CPT::Util::CRC64->new(); foreach my $file(@genbank_files){ my $seqio_object = Bio::SeqIO->new(-file => $file,-format=>'genbank'); while(my $seqobj = $seqio_object->next_seq){ my ( $fh, $path ) = tempfile('cds_export.XXXXXXXXX', UNLINK => $GLOBAL_UNLINK_VAR, DIR => $tempdir, SUFFIX => '.fa'); my @gi_array; foreach my $feat ( $seqobj->get_SeqFeatures ) { if($feat->primary_tag eq 'CDS'){ my $header = $bio->_getIdentifier($feat); # This ensures proteins have a file-specific ID appeneded to them. my $seq = $bio->translate( $bio->intelligent_get_seq($feat)); # Proteins come with translated stop codon $seq =~ s/\*//g; $seq =~ s/\+//g; $seq =~ s/#//g; $header .= "_" . $crc->crc64($seq); push @gi_array, $header; print $fh ">$header\n$seq\n"; } } $data{gbk}{$seqobj->display_id()}{'gi'} = \@gi_array; $data{gbk}{$seqobj->display_id()}{'fasta_location'} = $path; $data{gbk}{$seqobj->display_id()}{'gbk_location'} = $file; push(@{$data{file_list}}, $seqobj->display_id()); close $fh; } } use IPC::Run3; # Concatenate Fasta Files my @fasta_files; foreach(@{$data{file_list}}){ push(@fasta_files, $data{gbk}{$_}{fasta_location}); } my @command = ('cat', @fasta_files); my ($merged_fa_fh, $merged_fa_path) = tempfile('merged.XXXXXXXXX', UNLINK => 1, DIR => $tempdir, SUFFIX => '.fa'); my ($in, $out, $err); run3 \@command, \$in, \$out, \$err; if($err){ print STDERR $err; } print $merged_fa_fh $out; close($merged_fa_fh); # Create Blast Database my ($blastdb_fh, $blastdb_path) = tempfile('blastdb.XXXXXXXXX', UNLINK => 1, DIR => $tempdir); @command = ('makeblastdb', '-dbtype', 'prot', '-in', $merged_fa_path, '-out', $blastdb_path, ); my ($makeblast_in,$makeblast_out,$makeblast_err); run3 \@command, \$makeblast_in, \$makeblast_out, \$makeblast_err; # Blast files foreach(@{$data{file_list}}){ #push(@fasta_files, $data{gbk}{$_}{fasta_location}); my @blast_cmd = ('blastp', '-query', $data{gbk}{$_}{fasta_location}, '-out', $data{gbk}{$_}{fasta_location} . ".xml", '-outfmt', '5', '-db', $blastdb_path, ); my ($blast_in,$blast_out,$blast_err); run3 \@blast_cmd, \$blast_in, \$blast_out, \$blast_err; } my $value; my @data_tsv; foreach(@{$data{file_list}}){ #push(@fasta_files, $data{gbk}{$_}{fasta_location}); my $file = $data{gbk}{$_}{fasta_location}; my $in = new Bio::SearchIO( -format => 'blastxml', -tempfile => 1, -file => "$file.xml", ); while( my $result = $in->next_result ) { while( my $hit = $result->next_hit ) { while( my $hsp = $hit->next_hsp ) { my $Identity = $hsp->percent_identity/100 * $hsp->length('query'); my $IterationQueryLength = $result->query_length(); my $HitLength = $hit->length(); my $dice = (200*$Identity)/($HitLength + $IterationQueryLength); my $c = $result->query_description();#genome_a_header my $d = $hit->name();#genome_b_header # Skip self-self links next if($c eq $d); push (@data_tsv, [ $c, $d, $hsp->evalue(), $dice, ] ); }#close hsp }#close hit }#close result }#close for genomes $data{hit_table} = \@data_tsv; use CPT::OutputFiles; my $psmout = CPT::OutputFiles->new( name => 'cpt_psm_object', GGO => $ggo, ); my @val = $psmout->CRR(data => "none", extension => 'psm'); store \%data,$val[0]; =head1 NAME PSM Prep =head1 DESCRIPTION This tool takes in 2 or more GenBank files, blasts, and prepares data structures for use in the companion tool: PSM Plotter. Select as many (multi)-gbk files as you I<might> want to plot. Once this tool is done, you can select any subset of those to plot then. =cut