#!/usr/bin/perl #------------------------------------------------------------------------------- # Programmer: Adam M Phillippy, The Institute for Genomic Research # File: nucmer # Date: 04 / 09 / 03 # # Usage: # nucmer [options] # # Try 'nucmer -h' for more information. # # Purpose: To create alignments between two multi-FASTA inputs by using # the MUMmer matching and clustering algorithms. # #------------------------------------------------------------------------------- use lib "/home/psgendb/local/pkg/MUMmer3.22/scripts"; use Foundation; use File::Spec::Functions; use strict; my $AUX_BIN_DIR = "/home/psgendb/local/pkg/MUMmer3.22/aux_bin"; my $BIN_DIR = "/home/psgendb/local/pkg/MUMmer3.22"; my $SCRIPT_DIR = "/home/psgendb/local/pkg/MUMmer3.22/scripts"; my $VERSION_INFO = q~ NUCmer (NUCleotide MUMmer) version 3.07 ~; my $HELP_INFO = q~ USAGE: nucmer [options] DESCRIPTION: nucmer generates nucleotide alignments between two mutli-FASTA input files. The out.delta output file lists the distance between insertions and deletions that produce maximal scoring alignments between each sequence. The show-* utilities know how to read this format. MANDATORY: Reference Set the input reference multi-FASTA filename Query Set the input query multi-FASTA filename OPTIONS: --mum Use anchor matches that are unique in both the reference and query --mumcand Same as --mumreference --mumreference Use anchor matches that are unique in in the reference but not necessarily unique in the query (default behavior) --maxmatch Use all anchor matches regardless of their uniqueness -b|breaklen Set the distance an alignment extension will attempt to extend poor scoring regions before giving up (default 200) -c|mincluster Sets the minimum length of a cluster of matches (default 65) --[no]delta Toggle the creation of the delta file (default --delta) --depend Print the dependency information and exit -d|diagfactor Set the clustering diagonal difference separation factor (default 0.12) --[no]extend Toggle the cluster extension step (default --extend) -f --forward Use only the forward strand of the Query sequences -g|maxgap Set the maximum gap between two adjacent matches in a cluster (default 90) -h --help Display help information and exit -l|minmatch Set the minimum length of a single match (default 20) -o --coords Automatically generate the original NUCmer1.1 coords output file using the 'show-coords' program --[no]optimize Toggle alignment score optimization, i.e. if an alignment extension reaches the end of a sequence, it will backtrack to optimize the alignment score instead of terminating the alignment at the end of the sequence (default --optimize) -p|prefix Set the prefix of the output files (default "out") -r --reverse Use only the reverse complement of the Query sequences --[no]simplify Simplify alignments by removing shadowed clusters. Turn this option off if aligning a sequence to itself to look for repeats (default --simplify) -V --version Display the version information and exit ~; my $USAGE_INFO = q~ USAGE: nucmer [options] ~; my @DEPEND_INFO = ( "$BIN_DIR/mummer", "$BIN_DIR/mgaps", "$BIN_DIR/show-coords", "$AUX_BIN_DIR/postnuc", "$AUX_BIN_DIR/prenuc", "$SCRIPT_DIR/Foundation.pm" ); my %DEFAULT_PARAMETERS = ( "OUTPUT_PREFIX" => "out", # prefix for all output files "MATCH_ALGORITHM" => "-mumreference", # match finding algo switch "MATCH_DIRECTION" => "-b", # match direction switch "MIN_MATCH" => "20", # minimum match size "MAX_GAP" => "90", # maximum gap between matches "MIN_CLUSTER" => "65", # minimum cluster size "DIAG_FACTOR" => ".12", # diagonal difference fraction "BREAK_LEN" => "200", # extension break length "POST_SWITCHES" => "" # switches for the post processing ); sub main ( ) { my $tigr; # TIGR::Foundation object my @err; # Error variable my $ref_file; # path of the reference input file my $qry_file; # path of the query input file #-- The command line options for the various programs my $pfx = $DEFAULT_PARAMETERS { "OUTPUT_PREFIX" }; my $algo = $DEFAULT_PARAMETERS { "MATCH_ALGORITHM" }; my $mdir = $DEFAULT_PARAMETERS { "MATCH_DIRECTION" }; my $size = $DEFAULT_PARAMETERS { "MIN_MATCH" }; my $gap = $DEFAULT_PARAMETERS { "MAX_GAP" }; my $clus = $DEFAULT_PARAMETERS { "MIN_CLUSTER" }; my $diff = $DEFAULT_PARAMETERS { "DIAG_FACTOR" }; my $blen = $DEFAULT_PARAMETERS { "BREAK_LEN" }; my $psw = $DEFAULT_PARAMETERS { "POST_SWITCHES" }; my $fwd; # if true, use forward strand my $rev; # if true, use reverse strand my $maxmatch; # matching algorithm switches my $mumreference; my $mum; my $extend = 1; # if true, extend clusters my $delta = 1; # if true, create the delta file my $optimize = 1; # if true, optimize alignment scores my $simplify = 1; # if true, simplify shadowed alignments my $generate_coords; #-- Initialize TIGR::Foundation $tigr = new TIGR::Foundation; if ( !defined ($tigr) ) { print (STDERR "ERROR: TIGR::Foundation could not be initialized"); exit (1); } #-- Set help and usage information $tigr->setHelpInfo ($HELP_INFO); $tigr->setUsageInfo ($USAGE_INFO); $tigr->setVersionInfo ($VERSION_INFO); $tigr->addDependInfo (@DEPEND_INFO); #-- Get command line parameters $err[0] = $tigr->TIGR_GetOptions ( "maxmatch" => \$maxmatch, "mumcand" => \$mumreference, "mumreference" => \$mumreference, "mum" => \$mum, "b|breaklen=i" => \$blen, "c|mincluster=i" => \$clus, "delta!" => \$delta, "d|diagfactor=f" => \$diff, "extend!" => \$extend, "f|forward" => \$fwd, "g|maxgap=i" => \$gap, "l|minmatch=i" => \$size, "o|coords" => \$generate_coords, "optimize!" => \$optimize, "p|prefix=s" => \$pfx, "r|reverse" => \$rev, "simplify!" => \$simplify ); #-- Check if the parsing was successful if ( $err[0] == 0 || $#ARGV != 1 ) { $tigr->printUsageInfo( ); print (STDERR "Try '$0 -h' for more information.\n"); exit (1); } $ref_file = File::Spec->rel2abs ($ARGV[0]); $qry_file = File::Spec->rel2abs ($ARGV[1]); #-- Set up the program parameters if ( $fwd && $rev ) { $mdir = "-b"; } elsif ( $fwd ) { $mdir = ""; } elsif ( $rev ) { $mdir = "-r"; } if ( ! $extend ) { $psw .= "-e "; } if ( ! $delta ) { $psw .= "-d "; } if ( ! $optimize ) { $psw .= "-t "; } if ( ! $simplify ) { $psw .= "-s "; } undef (@err); $err[0] = 0; if ( $mum ) { $err[0] ++; $algo = "-mum"; } if ( $mumreference ) { $err[0] ++; $algo = "-mumreference"; } if ( $maxmatch ) { $err[0] ++; $algo = "-maxmatch"; } if ( $err[0] > 1 ) { $tigr->printUsageInfo( ); print (STDERR "ERROR: Multiple matching algorithms selected\n"); print (STDERR "Try '$0 -h' for more information.\n"); exit (1); } #-- Set up the program path names my $algo_path = "$BIN_DIR/mummer"; my $mgaps_path = "$BIN_DIR/mgaps"; my $prenuc_path = "$AUX_BIN_DIR/prenuc"; my $postnuc_path = "$AUX_BIN_DIR/postnuc"; my $showcoords_path = "$BIN_DIR/show-coords"; #-- Check that the files needed are all there and readable/writable { undef (@err); if ( !$tigr->isExecutableFile ($algo_path) ) { push (@err, $algo_path); } if ( !$tigr->isExecutableFile ($mgaps_path) ) { push (@err, $mgaps_path); } if ( !$tigr->isExecutableFile ($prenuc_path) ) { push (@err, $prenuc_path); } if ( !$tigr->isExecutableFile ($postnuc_path) ) { push (@err, $postnuc_path); } if ( !$tigr->isReadableFile ($ref_file) ) { push (@err, $ref_file); } if ( !$tigr->isReadableFile ($qry_file) ) { push (@err, $qry_file); } if ( !$tigr->isCreatableFile ("$pfx.ntref") ) { if ( !$tigr->isWritableFile ("$pfx.ntref") ) { push (@err, "$pfx.ntref"); } } if ( !$tigr->isCreatableFile ("$pfx.mgaps") ) { if ( !$tigr->isWritableFile ("$pfx.mgaps") ) { push (@err, "$pfx.mgaps"); } } if ( !$tigr->isCreatableFile ("$pfx.delta") ) { if ( !$tigr->isWritableFile ("$pfx.delta") ) { push (@err, "$pfx.delta"); } } if ( $generate_coords ) { if ( !$tigr->isExecutableFile ($showcoords_path) ) { push (@err, $showcoords_path); } if ( !$tigr->isCreatableFile ("$pfx.coords") ) { if ( !$tigr->isWritableFile ("$pfx.coords") ) { push (@err, "$pfx.coords"); } } } #-- If 1 or more files could not be processed, terminate script if ( $#err >= 0 ) { $tigr->logError ("ERROR: The following critical files could not be used", 1); while ( $#err >= 0 ) { $tigr->logError (pop(@err), 1); } $tigr->logError ("Check your paths and file permissions and try again", 1); $tigr->bail( ); } } #-- Run prenuc and assert return value is zero print (STDERR "1: PREPARING DATA\n"); $err[0] = $tigr->runCommand ("$prenuc_path $ref_file > $pfx.ntref"); if ( $err[0] != 0 ) { $tigr->bail ("ERROR: prenuc returned non-zero\n"); } #-- Run mummer | mgaps and assert return value is zero print (STDERR "2,3: RUNNING mummer AND CREATING CLUSTERS\n"); open(ALGO_PIPE, "$algo_path $algo $mdir -l $size -n $pfx.ntref $qry_file |") or $tigr->bail ("ERROR: could not open $algo_path output pipe $!"); open(CLUS_PIPE, "| $mgaps_path -l $clus -s $gap -f $diff > $pfx.mgaps") or $tigr->bail ("ERROR: could not open $mgaps_path input pipe $!"); while ( ) { print CLUS_PIPE or $tigr->bail ("ERROR: could not write to $mgaps_path pipe $!"); } $err[0] = close(ALGO_PIPE); $err[1] = close(CLUS_PIPE); if ( $err[0] == 0 || $err[1] == 0 ) { $tigr->bail ("ERROR: mummer and/or mgaps returned non-zero\n"); } #-- Run postnuc and assert return value is zero print (STDERR "4: FINISHING DATA\n"); $err[0] = $tigr->runCommand ("$postnuc_path $psw -b $blen $ref_file $qry_file $pfx < $pfx.mgaps"); if ( $err[0] != 0 ) { $tigr->bail ("ERROR: postnuc returned non-zero\n"); } #-- If the -o flag was set, run show-coords using NUCmer1.1 settings if ( $generate_coords ) { print (STDERR "5: GENERATING COORDS FILE\n"); $err[0] = $tigr->runCommand ("$showcoords_path -r $pfx.delta > $pfx.coords"); if ( $err[0] != 0 ) { $tigr->bail ("ERROR: show-coords returned non-zero\n"); } } #-- Remove the temporary output $err[0] = unlink ("$pfx.ntref", "$pfx.mgaps"); if ( $err[0] != 2 ) { $tigr->logError ("WARNING: there was a problem deleting". " the temporary output files", 1); } #-- Return success return (0); } exit ( main ( ) ); #-- END OF SCRIPT