# $Id: SeqWords.pm 16123 2009-09-17 12:57:27Z cjfields $ #--------------------------------------------------------------------------- # PACKAGE : Bio::Tools::SeqWords # PURPOSE : To count n-mers in any sequence of characters # AUTHOR : Derek Gatherer (d.gatherer@vir.gla.ac.uk) # SOURCE : # CREATED : 21st March 2000 # MODIFIED : 11th November 2003 (DG - new method, count_overlap_words) # LICENCE : You may distribute this module under the same terms # : as the rest of BioPerl. #--------------------------------------------------------------------------- =head1 NAME Bio::Tools::SeqWords - Object holding n-mer statistics for a sequence =head1 SYNOPSIS # Create the SeqWords object, e.g.: my $inputstream = Bio::SeqIO->new(-file => "seqfile", -format => 'Fasta'); my $seqobj = $inputstream->next_seq(); my $seq_word = Bio::Tools::SeqWords->new(-seq => $seqobj); # Or: my $seqobj = Bio::PrimarySeq->new(-seq => "agggtttccc", -alphabet => 'dna', -id => 'test'); my $seq_word = Bio::Tools::SeqWords->new(-seq => $seqobj); # obtain a hash of word counts, eg: my $hash_ref = $seq_stats->count_words($word_length); # display hash table, eg: my %hash = %$hash_ref; foreach my $key(sort keys %hash) { print "\n$key\t$hash{$key}"; } # Or: my $hash_ref = Bio::Tools::SeqWords->count_words($seqobj,$word_length); =head1 DESCRIPTION L is a featherweight object for the calculation of n-mer word occurrences in a single sequence. It is envisaged that the object will be useful for construction of scripts which use n-mer word tables as the raw material for statistical calculations; for instance, hexamer frequency for the calculation of coding protential, or the calculation of periodicity in repetitive DNA. Triplet frequency is already handled by L (author: Peter Schattner). There are a few possible applications for protein, e.g. hypothesised amino acid 7-mers in heat shock proteins, or proteins with multiple simple motifs. Sometimes these protein periodicities are best seen when the amino acid alphabet is truncated, e.g. Shulman alphabet. Since there are quite a few of these shortened alphabets, this module does not specify any particular alphabet. See Synopsis above for object creation code. =head2 Rationale Take a sequence object and create an object for the purposes of holding n-mer word statistics about that sequence. The sequence can be nucleic acid or protein. In count_words() the words are counted in a non-overlapping manner, ie. in the style of a codon table, but with any word length. In count_overlap_words() the words are counted in an overlapping manner. For counts on opposite strand (DNA/RNA), a reverse complement method should be performed, and then the count repeated. =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 Derek Gatherer, in the loosest sense of the word 'author'. The general shape of the module is lifted directly from the SeqStat module of Peter Schattner. The central subroutine to count the words is adapted from original code provided by Dave Shivak, in response to a query on the bioperl mailing list. At least 2 other people provided alternative means (equally good but not used in the end) of performing the same calculation. Thanks to all for your assistance. =head1 CONTRIBUTORS Jason Stajich, jason-at-bioperl.org =head1 APPENDIX The rest of the documentation details each of the object methods. Internal methods are usually preceded with a _ =cut package Bio::Tools::SeqWords; use strict; use base qw(Bio::Root::Root); sub new { my($class,@args) = @_; # our new standard way of instantiation my $self = $class->SUPER::new(@args); my ($seqobj) = $self->_rearrange([qw(SEQ)],@args); if((! defined($seqobj)) && @args && ref($args[0])) { # parameter not passed as named parameter? $seqobj = $args[0]; } if(! $seqobj->isa("Bio::PrimarySeqI")) { $self->throw(ref($self) . " works only on PrimarySeqI objects\n"); } $self->{'_seqref'} = $seqobj; return $self; } =head2 count_words Title : count_words Usage : $word_count = $seq_stats->count_words($word_length) or $word_count = $seq_stats->Bio::Tools::SeqWords->($seqobj,$word_length); Function: Counts non-overlapping words within a string, any alphabet is used Example : a sequence ACCGTCCGT, counted at word length 4, will give the hash {ACCG => 1, TCCG => 1} Returns : Reference to a hash in which keys are words (any length) of the alphabet used and values are number of occurrences of the word in the sequence. Args : Word length as scalar and, reference to sequence object if required Throws an exception word length is not a positive integer or if word length is longer than the sequence. =cut sub count_words { my ($self,$seqobj,$word_length) = @_; # check how we were called, and if necessary rearrange arguments if(ref($seqobj)) { # call as SeqWords->count_words($seq, $wordlen) if(! $seqobj->isa("Bio::PrimarySeqI")) { $self->throw("SeqWords works only on PrimarySeqI objects\n"); } } else { # call as $obj->count_words($wordlen) $word_length = $seqobj; $seqobj = undef; } if(! defined($seqobj)){ $seqobj = $self->{'_seqref'}; } if($word_length eq "" || $word_length =~ /[a-z]/i){ $self->throw("SeqWords cannot accept non-numeric characters". " or a null value in the \$word_length variable\n"); }elsif ($word_length <1 || ($word_length - int($word_length)) >0){ $self->throw("SeqWords requires the word length to be a ". "positive integer\n"); } my $seqstring = uc $seqobj->seq(); if($word_length > length($seqstring)){ $self->throw("die in _count, \$word_length is bigger ". "than sequence length\n"); } my $type = "non-overlap"; my $words = _count($seqobj, $word_length, $type); return $words; # ref. to a hash } =head2 count_overlap_words Title : count_overlap_words Usage : $word_count = $word_obj->count_overlap_words($word_length); Function: Counts overlapping words within a string, any alphabet is used Example : A sequence ACCAACCA, counted at word length 4, will give the hash {ACCA=>2, CCAA=>1, CAAC=>1, AACC=>1} Returns : Reference to a hash in which keys are words (any length) of the alphabet used and values are number of occurrences of the word in the sequence. Args : Word length as scalar Throws an exception if word length is not a positive integer or if word length is longer than the sequence. =cut sub count_overlap_words { my ($self,$seqobj,$word_length) = @_; # check how we were called, and if necessary rearrange arguments if(ref($seqobj)){ # call as SeqWords->count_words($seq, $wordlen) if(! $seqobj->isa("Bio::PrimarySeqI")){ $self->throw("SeqWords works only on PrimarySeqI objects\n"); } }else{ # call as $obj->count_words($wordlen) $word_length = $seqobj; $seqobj = undef; } if(! defined($seqobj)) { $seqobj = $self->{'_seqref'}; } my $seqstring = uc $seqobj->seq(); if($word_length > length($seqstring)){ $self->throw("die in _count, \$word_length is bigger ". "than sequence length\n"); } my $type = "overlap"; my $words = _count($seqobj, $word_length, $type); return $words; # ref. to a hash } # the actual counting routine # used by both count_words and count_overlap_words sub _count { my ($seqobj, $word_length, $type) = @_; my %codon = (); # now the real business # JS - remove DNA assumption my $seqstring = uc $seqobj->seq(); if($type eq "non-overlap") { while($seqstring =~ /((\w){$word_length})/gim){ $codon{uc($1)}++; } } elsif($type eq "overlap"){ my $seqlen = $seqobj->length(); # measure length for (my $frame = 1; $frame <= $word_length; $frame++) { # run through frames my $seqstring = uc($seqobj->subseq($frame,$seqlen)); # take the relevant substring while($seqstring =~ /((\w){$word_length})/gim){ $codon{uc($1)}++; # keep adding to hash } } } else { Bio::Root::Root->throw("\nSomething badly wrong here. \$type: $type can only be overlap or non-overlap"); } return \%codon; } 1;