# $Id: Perl.pm 16123 2009-09-17 12:57:27Z cjfields $ # # BioPerl module for Bio::Perl # # Please direct questions and support issues to # # Cared for by Ewan Birney # # Copyright Ewan Birney # # You may distribute this module under the same terms as perl itself # POD documentation - main docs before the code =head1 NAME Bio::Perl - Functional access to BioPerl for people who don't know objects =head1 SYNOPSIS use Bio::Perl; # will guess file format from extension $seq_object = read_sequence($filename); # forces genbank format $seq_object = read_sequence($filename,'genbank'); # reads an array of sequences @seq_object_array = read_all_sequences($filename,'fasta'); # sequences are Bio::Seq objects, so the following methods work # for more info see Bio::Seq, or do 'perldoc Bio/Seq.pm' print "Sequence name is ",$seq_object->display_id,"\n"; print "Sequence acc is ",$seq_object->accession_number,"\n"; print "First 5 bases is ",$seq_object->subseq(1,5),"\n"; # get the whole sequence as a single string $sequence_as_a_string = $seq_object->seq(); # writing sequences write_sequence(">$filename",'genbank',$seq_object); write_sequence(">$filename",'genbank',@seq_object_array); # making a new sequence from just a string $seq_object = new_sequence("ATTGGTTTGGGGACCCAATTTGTGTGTTATATGTA", "myname","AL12232"); # getting a sequence from a database (assumes internet connection) $seq_object = get_sequence('swissprot',"ROA1_HUMAN"); $seq_object = get_sequence('embl',"AI129902"); $seq_object = get_sequence('genbank',"AI129902"); # BLAST a sequence (assummes an internet connection) $blast_report = blast_sequence($seq_object); write_blast(">blast.out",$blast_report); =head1 DESCRIPTION Easy first time access to BioPerl via functions. =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 =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 - Ewan Birney Email birney@ebi.ac.uk =head1 APPENDIX The rest of the documentation details each of the object methods. Internal methods are usually preceded with a _ =cut #' # Let the code begin... package Bio::Perl; use vars qw(@EXPORT @EXPORT_OK $DBOKAY); use strict; use Carp; use Bio::SeqIO; use Bio::Seq; use Bio::Root::Version '$VERSION'; BEGIN { eval { require Bio::DB::EMBL; require Bio::DB::GenBank; require Bio::DB::SwissProt; require Bio::DB::RefSeq; require Bio::DB::GenPept; }; if( $@ ) { $DBOKAY = 0; } else { $DBOKAY = 1; } } use base qw(Exporter); @EXPORT = qw(read_sequence read_all_sequences write_sequence new_sequence get_sequence translate translate_as_string reverse_complement revcom revcom_as_string reverse_complement_as_string blast_sequence write_blast); @EXPORT_OK = @EXPORT; =head2 read_sequence Title : read_sequence Usage : $seq = read_sequence('sequences.fa') $seq = read_sequence($filename,'genbank'); # pipes are fine $seq = read_sequence("my_fetching_program $id |",'fasta'); Function: Reads the top sequence from the file. If no format is given, it will try to guess the format from the filename. If a format is given, it forces that format. The filename can be any valid perl open() string - in particular, you can put in pipes Returns : A Bio::Seq object. A quick synopsis: $seq_object->display_id - name of the sequence $seq_object->seq - sequence as a string Args : Two strings, first the filename - any Perl open() string is ok Second string is the format, which is optional For more information on Seq objects see L. =cut sub read_sequence{ my ($filename,$format) = @_; if( !defined $filename ) { confess "read_sequence($filename) - usage incorrect"; } my $seqio; if( defined $format ) { $seqio = Bio::SeqIO->new( '-file' => $filename, '-format' => $format); } else { $seqio = Bio::SeqIO->new( '-file' => $filename); } my $seq = $seqio->next_seq(); return $seq; } =head2 read_all_sequences Title : read_all_sequences Usage : @seq_object_array = read_all_sequences($filename); @seq_object_array = read_all_sequences($filename,'genbank'); Function: Just as the function above, but reads all the sequences in the file and loads them into an array. For very large files, you will run out of memory. When this happens, you've got to use the SeqIO system directly (this is not so hard! Don't worry about it!). Returns : array of Bio::Seq objects Args : two strings, first the filename (any open() string is ok) second the format (which is optional) See L and L for more information =cut sub read_all_sequences{ my ($filename,$format) = @_; if( !defined $filename ) { confess "read_all_sequences($filename) - usage incorrect"; } my $seqio; if( defined $format ) { $seqio = Bio::SeqIO->new( '-file' => $filename, '-format' => $format); } else { $seqio = Bio::SeqIO->new( '-file' => $filename); } my @seq_array; while( my $seq = $seqio->next_seq() ) { push(@seq_array,$seq); } return @seq_array; } =head2 write_sequence Title : write_sequence Usage : write_sequence(">new_file.gb",'genbank',$seq) write_sequence(">new_file.gb",'genbank',@array_of_sequence_objects) Function: writes sequences in the specified format Returns : true Args : filename as a string, must provide an open() output file format as a string one or more sequence objects =cut sub write_sequence{ my ($filename,$format,@sequence_objects) = @_; if( scalar(@sequence_objects) == 0 ) { confess("write_sequence(filename,format,sequence_object)"); } my $error = 0; my $seqname = "sequence1"; # catch users who haven't passed us a filename we can open if( $filename !~ /^\>/ && $filename !~ /^|/ ) { $filename = ">".$filename; } my $seqio = Bio::SeqIO->new('-file' => $filename, '-format' => $format); foreach my $seq ( @sequence_objects ) { my $seq_obj; if( !ref $seq ) { if( length $seq > 50 ) { # odds are this is a sequence as a string, and someone has not figured out # how to make objects. Warn him/her and then make a sequence object # from this if( $error == 0 ) { carp("WARNING: You have put in a long string into write_sequence.\nI suspect this means that this is the actual sequence\nIn the future try the\n new_sequence method of this module to make a new sequence object.\nDoing this for you here\n"); $error = 1; } $seq_obj = new_sequence($seq,$seqname); $seqname++; } else { confess("You have a non object [$seq] passed to write_sequence. It maybe that you want to use new_sequence to make this string into a sequence object?"); } } else { if( !$seq->isa("Bio::SeqI") ) { confess("object [$seq] is not a Bio::Seq object; can't write it out"); } $seq_obj = $seq; } # finally... we get to write out the sequence! $seqio->write_seq($seq_obj); } 1; } =head2 new_sequence Title : new_sequence Usage : $seq_obj = new_sequence("GATTACA", "kino-enzyme"); Function: Construct a sequency object from sequence string Returns : A Bio::Seq object Args : sequence string name string (optional, default "no-name-for-sequence") accession - accession number (optional, no default) =cut sub new_sequence{ my ($seq,$name,$accession) = @_; if( !defined $seq ) { confess("new_sequence(sequence_as_string) usage"); } $name ||= "no-name-for-sequence"; my $seq_object = Bio::Seq->new( -seq => $seq, -id => $name); $accession && $seq_object->accession_number($accession); return $seq_object; } =head2 blast_sequence Title : blast_sequence Usage : $blast_result = blast_sequence($seq) $blast_result = blast_sequence('MFVEGGTFASEDDDSASAEDE'); Function: If the computer has Internet accessibility, blasts the sequence using the NCBI BLAST server against nrdb. It chooses the flavour of BLAST on the basis of the sequence. This function uses Bio::Tools::Run::RemoteBlast, which itself use Bio::SearchIO - as soon as you want to know more, check out these modules Returns : Bio::Search::Result::GenericResult.pm Args : Either a string of protein letters or nucleotides, or a Bio::Seq object =cut sub blast_sequence { my ($seq,$verbose) = shift; if( !defined $verbose ) { $verbose = 1; } if( !ref $seq ) { $seq = Bio::Seq->new( -seq => $seq, -id => 'blast-sequence-temp-id'); } elsif ( !$seq->isa('Bio::PrimarySeqI') ) { croak("[$seq] is an object, but not a Bio::Seq object, cannot be blasted"); } require Bio::Tools::Run::RemoteBlast; my $prog = 'blastp'; my $e_val= '1e-10'; my @params = ( '-prog' => $prog, '-expect' => $e_val, '-readmethod' => 'SearchIO' ); my $factory = Bio::Tools::Run::RemoteBlast->new(@params); my $r = $factory->submit_blast($seq); if( $verbose ) { print STDERR "Submitted Blast for [".$seq->id."] "; } sleep 5; my $result; LOOP : while( my @rids = $factory->each_rid) { foreach my $rid ( @rids ) { my $rc = $factory->retrieve_blast($rid); if( !ref($rc) ) { if( $rc < 0 ) { $factory->remove_rid($rid); } if( $verbose ) { print STDERR "."; } sleep 10; } else { $result = $rc->next_result(); $factory->remove_rid($rid); last LOOP; } } } if( $verbose ) { print STDERR "\n"; } return $result; } =head2 write_blast Title : write_blast Usage : write_blast($filename,$blast_report); Function: Writes a BLAST result object (or more formally a SearchIO result object) out to a filename in BLAST-like format Returns : none Args : filename as a string Bio::SearchIO::Results object =cut sub write_blast { my ($filename,$blast) = @_; if( $filename !~ /^\>/ && $filename !~ /^|/ ) { $filename = ">".$filename; } my $output = Bio::SearchIO->new( -output_format => 'blast', -file => $filename); $output->write_result($blast); } =head2 get_sequence Title : get_sequence Usage : $seq_object = get_sequence('swiss',"ROA1_HUMAN"); Function: If the computer has Internet access this method gets the sequence from Internet accessible databases. Currently this supports Swissprot ('swiss'), EMBL ('embl'), GenBank ('genbank'), GenPept ('genpept'), and RefSeq ('refseq'). Swissprot and EMBL are more robust than GenBank fetching. If the user is trying to retrieve a RefSeq entry from GenBank/EMBL, the query is silently redirected. Returns : A Bio::Seq object Args : database type - one of swiss, embl, genbank, genpept, or refseq =cut my $genbank_db = undef; my $genpept_db = undef; my $embl_db = undef; my $swiss_db = undef; my $refseq_db = undef; sub get_sequence{ my ($db_type,$identifier) = @_; if( ! $DBOKAY ) { confess ("Your system does not have one of LWP, HTTP::Request::Common, IO::String installed so the DB retrieval method is not available. \nFull error message is:\n $!\n"); return; } $db_type = lc($db_type); my $db; if( $db_type =~ /genbank/ ) { if( !defined $genbank_db ) { $genbank_db = Bio::DB::GenBank->new(); } $db = $genbank_db; } if( $db_type =~ /genpept/ ) { if( !defined $genpept_db ) { $genpept_db = Bio::DB::GenPept->new(); } $db = $genpept_db; } if( $db_type =~ /swiss/ ) { if( !defined $swiss_db ) { $swiss_db = Bio::DB::SwissProt->new(); } $db = $swiss_db; } if( $db_type =~ /embl/ ) { if( !defined $embl_db ) { $embl_db = Bio::DB::EMBL->new(); } $db = $embl_db; } if( $db_type =~ /refseq/ or ($db_type !~ /swiss/ and $identifier =~ /^\s*N\S+_/)) { if( !defined $refseq_db ) { $refseq_db = Bio::DB::RefSeq->new(); } $db = $refseq_db; } my $seq; if( $identifier =~ /^\w+\d+$/ ) { $seq = $db->get_Seq_by_acc($identifier); } else { $seq = $db->get_Seq_by_id($identifier); } return $seq; } =head2 translate Title : translate Usage : $seqobj = translate($seq_or_string_scalar) Function: translates a DNA sequence object OR just a plain string of DNA to amino acids Returns : A Bio::Seq object Args : Either a sequence object or a string of just DNA sequence characters =cut sub translate { my ($scalar) = shift; my $obj; if( ref $scalar ) { if( !$scalar->isa("Bio::PrimarySeqI") ) { confess("Expecting a sequence object not a $scalar"); } else { $obj= $scalar; } } else { # check this looks vaguely like DNA my $n = ( $scalar =~ tr/ATGCNatgcn/ATGCNatgcn/ ); if( $n < length($scalar) * 0.85 ) { confess("Sequence [$scalar] is less than 85% ATGCN, which doesn't look very DNA to me"); } $obj = Bio::PrimarySeq->new(-id => 'internalbioperlseq',-seq => $scalar); } return $obj->translate(); } =head2 translate_as_string Title : translate_as_string Usage : $seqstring = translate_as_string($seq_or_string_scalar) Function: translates a DNA sequence object OR just a plain string of DNA to amino acids Returns : A string of just amino acids Args : Either a sequence object or a string of just DNA sequence characters =cut sub translate_as_string { my ($scalar) = shift; my $obj = Bio::Perl::translate($scalar); return $obj->seq; } =head2 reverse_complement Title : reverse_complement Usage : $seqobj = reverse_complement($seq_or_string_scalar) Function: reverse complements a string or sequence argument producing a Bio::Seq - if you want a string, you can use reverse_complement_as_string Returns : A Bio::Seq object Args : Either a sequence object or a string of just DNA sequence characters =cut sub reverse_complement { my ($scalar) = shift; my $obj; if( ref $scalar ) { if( !$scalar->isa("Bio::PrimarySeqI") ) { confess("Expecting a sequence object not a $scalar"); } else { $obj= $scalar; } } else { # check this looks vaguely like DNA my $n = ( $scalar =~ tr/ATGCNatgc/ATGCNatgcn/ ); if( $n < length($scalar) * 0.85 ) { confess("Sequence [$scalar] is less than 85% ATGCN, which doesn't look very DNA to me"); } $obj = Bio::PrimarySeq->new(-id => 'internalbioperlseq',-seq => $scalar); } return $obj->revcom(); } =head2 revcom Title : revcom Usage : $seqobj = revcom($seq_or_string_scalar) Function: reverse complements a string or sequence argument producing a Bio::Seq - if you want a string, you can use reverse_complement_as_string This is an alias for reverse_complement Returns : A Bio::Seq object Args : Either a sequence object or a string of just DNA sequence characters =cut sub revcom { return &Bio::Perl::reverse_complement(@_); } =head2 reverse_complement_as_string Title : reverse_complement_as_string Usage : $string = reverse_complement_as_string($seq_or_string_scalar) Function: reverse complements a string or sequence argument producing a string Returns : A string of DNA letters Args : Either a sequence object or a string of just DNA sequence characters =cut sub reverse_complement_as_string { my ($scalar) = shift; my $obj = &Bio::Perl::reverse_complement($scalar); return $obj->seq; } =head2 revcom_as_string Title : revcom_as_string Usage : $string = revcom_as_string($seq_or_string_scalar) Function: reverse complements a string or sequence argument producing a string Returns : A string of DNA letters Args : Either a sequence object or a string of just DNA sequence characters =cut sub revcom_as_string { my ($scalar) = shift; my $obj = &Bio::Perl::reverse_complement($scalar); return $obj->seq; } 1;