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#!/usr/bin/env perl
use FindBin;
use strict;
use warnings;
use Getopt::Long qw(:config no_ignore_case bundling pass_through);
use Data::Dumper;
use List::Util qw (min max);
use File::Basename;
use lib ("$FindBin::RealBin/PerlLib");
use POSIX qw(ceil);
use Gene_obj;
use Nuc_translator;
use Fasta_reader;
use Longest_orf;
my $UTIL_DIR = "$FindBin::RealBin/util";
my $help;
my $workdir;
my $verbose;
my ($reuse,$pfam_out);
my $CPU = 2;
my $usage = <<_EOH_;
######################################## Options ###################################################################################
#
###############
# ** Required:
###############
# --pep <string> peptide files
#
# --pfam_db <string> /path/to/pfam_db.hmm to search
# using hmmscan (which should be accessible via your PATH setting)
#
################
# ** Optional:
################
#
#
# --reuse If this option is given, any existing files are not overwritten but reused
#
#
# --pfam_out|o <string> You can also pre-run the pfam searches if --reuse is set. In that case,
# --pfam_out is the output of hhmscan --domtblout using --noali --cut_nc --acc --notextw
#
# --prepare_pfam Prepare data for PFAM search and then quit (for running PFAM on HPC/computing cluster
# with or without MPI )
#
# --workdir Force temporary output directory to this directory (e.g. if --reuse is needed)
#
#
# -h print this option menu and quit
# -v verbose
#
# --CPU <int> number of threads to use; (default: 2)
#
# --MPI use MPI (via ffindex_apply_mpi)
#
# --quiet send stderr to /dev/null
#
#
####################################################################################################################################
_EOH_
;
my $MPI_DEBUG = 0;
my $pep_file;
my $search_pfam;
my $prepare_pfam_only = 0;
my $MPI = 0;
&GetOptions( 'pep=s' => \$pep_file,
'h' => \$help,
'v' => \$verbose,
'CPU=i' => \$CPU,
'pfam_db=s' => \$search_pfam,
'reuse' => \$reuse,
'workdir:s' => \$workdir,
'pfam_out|o=s' => \$pfam_out,
'prepare_pfam' => \$prepare_pfam_only,
'debug' => \$MPI_DEBUG,
'MPI' => \$MPI,
);
if ($help) {
die $usage;
}
if (@ARGV) {
die "Error, don't understand options: @ARGV";
}
$|++;
our $SEE = $verbose;
unless ($pep_file && $search_pfam) {
die "$usage\n";
}
&check_for_pfam_execs if ($search_pfam);
$workdir = "transdecoder.tmp.$$" unless $workdir;
mkdir($workdir) unless -d $workdir;
die "Error, cannot mkdir $workdir" unless -d $workdir;
unless ($pfam_out) {
$pfam_out = basename($pep_file) . ".transdecoder.pfam.dat";
}
main: {
my $parafly_cmd_file = &multithread($pep_file);
my $parafly_cmd = "ParaFly -CPU $CPU -c $parafly_cmd_file --failed $parafly_cmd_file.failed ";
if (!$MPI_DEBUG) {
$parafly_cmd .= " -v ";
}
else {
# a little more verbose
$parafly_cmd .= " -vv ";
}
if ($prepare_pfam_only) {
print "We have prepared the $parafly_cmd_file command file for you to run hmmscan separately (e.g. on a cluster).\n";
print "Example for a single node with $CPU CPUs :\n\t$parafly_cmd\n\n";
print "After your PFAM searches are complete, then concatanate all the out.db files using this command into $pep_file.pfam.out.\n";
print "cat $workdir/*.out.db"
. '|tr -d \'\000\' '
. "|grep -v '^#' > $pep_file.pfam.out\n\n";
print "Then in order to restart transdecoder use the following command (along with any other options you want):\n";
print "\t$0 --pep $pep_file --search_pfam $search_pfam --pfam_out $pep_file.pfam.out --reuse --workdir $workdir\n\n";
exit(0);
}
print "Processing with PFAM HMM searches...\n";
unless ($reuse && -s $pfam_out){
&process_cmd("$parafly_cmd");
if (-s "$parafly_cmd_file.failed"){
die "Some sequences failed to be searched against PFAM. Please resolve the situation (see $parafly_cmd_file), delete $parafly_cmd_file and use --rerun and --workdir $workdir to resume.\n";
}
if ($MPI) {
&process_cmd("cat $workdir/*.out.db" . '|tr -d "\000" | grep -v "^#" > ' . $pfam_out);
&process_cmd("find $workdir -name '*.out.db' -delete");
&process_cmd("find $workdir -name '*.out.db.idx' -delete");
&process_cmd("find $workdir -name 'parafly.sh*' -delete");
unless (-s $pfam_out) {
die "Error, pfam results were not properly concatenated into file: $pfam_out";
}
}
else {
## Parse regular pfam table output files:
&process_cmd("find $workdir/ -name '*.fa.domtbl' -exec cat {} \\\; | egrep -v '^\#' > $pfam_out");
}
}
print STDERR "PFAM SEARCH DONE.\n\n";
exit(0);
}
####
sub process_cmd {
my ($cmd) = @_;
print "CMD: $cmd\n";
my $ret = system($cmd);
if ($ret) {
die "Error, cmd: $cmd died with ret $ret";
}
return;
}
sub index_fasta(){
# this where ffindex would really help
my $fasta_file = shift;
&process_cmd("$UTIL_DIR/bin/ffindex_from_fasta -s $fasta_file.db $fasta_file.db.idx $fasta_file") unless -s "$fasta_file.db";
my $sequence_number = `wc -l < $fasta_file.db.idx`;
chomp($sequence_number);
die unless $sequence_number;
return $sequence_number;
}
# MPIrun might require expert users here.... so just use parafly and be done with it (experts can run mpirun directly)
sub multithread(){
my ($protein_file) = @_;
print STDERR "Partitioning fasta file $protein_file\n";
my @fasta_files = &partition_transcript_db($protein_file);
my $cmd_file = "$workdir/parafly.sh";
unlink($cmd_file.'.completed');
unlink($cmd_file.'.failed');
open(OUT,">$cmd_file") || die;
foreach my $fasta_file (@fasta_files){
if ($MPI) {
my $cmd = "hmmscan -o /dev/null --cpu 1 --noali --cut_nc --acc --notextw --domtblout /dev/stdout $search_pfam - ";
my $sequence_number = &index_fasta($fasta_file);
my $pfam_out = "$fasta_file.out";
my $ffidx = "$UTIL_DIR/bin/ffindex_apply_mpi -d $pfam_out.db -i $pfam_out.idx $fasta_file.db $fasta_file.db.idx -- ";
unlink($pfam_out);
unlink($pfam_out.'.db');
unlink($pfam_out.'.idx');
if (!$MPI_DEBUG) {
$ffidx .= " 2>/dev/null";
}
print OUT $ffidx . " $cmd \n";
}
else {
## use ParaFly / hmmscan w/o ffindex
my $cmd = "hmmscan -o /dev/null --cpu 1 --noali --cut_nc --acc --notextw --domtblout $fasta_file.domtbl $search_pfam $fasta_file ";
print OUT "$cmd\n";
}
}
close OUT;
return "$cmd_file";
}
sub partition_transcript_db {
my $transcript_db = shift;
my $number_of_peps = `grep '>' $transcript_db | wc -l `;
chomp $number_of_peps;
my $seqs_per_partition = ceil($number_of_peps/$CPU);
$seqs_per_partition = 1 if $seqs_per_partition < 1;
$seqs_per_partition = $seqs_per_partition < 5000 ? $seqs_per_partition : 5000 ;
my @files;
my $fasta_reader = new Fasta_reader($transcript_db);
my $partition_counter = 0;
my $counter = 0;
my $ofh;
while (my $seq_obj = $fasta_reader->next()) {
my $fasta_entry = $seq_obj->get_FASTA_format();
$fasta_entry=~s/[\*\s]+$//; #strip stop codon/empty space
$fasta_entry.="\n";
if ($counter % $seqs_per_partition == 0) {
close $ofh if $ofh;
$partition_counter++;
my $outfile = "$workdir/partition.$counter.fa";
open ($ofh, ">$outfile") or die "Error, cannot write to outfile: $outfile";
push (@files, $outfile);
}
print $ofh $fasta_entry;
$counter++;
}
close $ofh if $ofh;
return(@files);
}
sub check_for_pfam_execs(){
$ENV{PATH} .= ":$UTIL_DIR/bin"; # now can find 3rd party tools in PATH setting
$ENV{LD_LIBRARY_PATH} .= ":$UTIL_DIR/lib64/";
die "Error, cannot locate pfam database at: $search_pfam" unless (-s $search_pfam);
my @utils = qw(hmmscan ParaFly);
if ($MPI) {
push (@utils, qw(ffindex_apply_mpi ffindex_from_fasta));
}
my @programs = &check_program(@utils);
}
sub check_program() {
my @paths;
foreach my $prog (@_) {
my $path = `which $prog`;
die "Error, path to a required program ($prog) cannot be found\n\n"
unless $path =~ /^\//;
chomp($path);
$path = readlink($path) if -l $path;
push( @paths, $path );
}
return @paths;
}
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