# $Id: RemoteBlast.pm 16123 2009-09-17 12:57:27Z cjfields $ # # BioPerl module for Bio::Tools::Run::RemoteBlast # # Please direct questions and support issues to # # FORMERLY Cared for by Jason Stajich, Mat Wiepert # # Somewhat cared for by Roger Hall, Chris Fields (when they have time) # # Copyright Jason Stajich, Bioperl # # You may distribute this module under the same terms as perl itself # POD documentation - main docs before the code =head1 NAME Bio::Tools::Run::RemoteBlast - Object for remote execution of the NCBI Blast via HTTP =head1 SYNOPSIS #Remote-blast "factory object" creation and blast-parameter initialization use Bio::Tools::Run::RemoteBlast; use strict; my $prog = 'blastp'; my $db = 'swissprot'; my $e_val= '1e-10'; my @params = ( '-prog' => $prog, '-data' => $db, '-expect' => $e_val, '-readmethod' => 'SearchIO' ); my $factory = Bio::Tools::Run::RemoteBlast->new(@params); #change a query paramter $Bio::Tools::Run::RemoteBlast::HEADER{'ENTREZ_QUERY'} = 'Homo sapiens [ORGN]'; #change a retrieval parameter $Bio::Tools::Run::RemoteBlast::RETRIEVALHEADER{'DESCRIPTIONS'} = 1000; #remove a parameter delete $Bio::Tools::Run::RemoteBlast::HEADER{'FILTER'}; #$v is just to turn on and off the messages my $v = 1; my $str = Bio::SeqIO->new(-file=>'amino.fa' , -format => 'fasta' ); while (my $input = $str->next_seq()){ #Blast a sequence against a database: #Alternatively, you could pass in a file with many #sequences rather than loop through sequence one at a time #Remove the loop starting 'while (my $input = $str->next_seq())' #and swap the two lines below for an example of that. my $r = $factory->submit_blast($input); #my $r = $factory->submit_blast('amino.fa'); print STDERR "waiting..." if( $v > 0 ); 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); } print STDERR "." if ( $v > 0 ); sleep 5; } else { my $result = $rc->next_result(); #save the output my $filename = $result->query_name()."\.out"; $factory->save_output($filename); $factory->remove_rid($rid); print "\nQuery Name: ", $result->query_name(), "\n"; while ( my $hit = $result->next_hit ) { next unless ( $v > 0); print "\thit name is ", $hit->name, "\n"; while( my $hsp = $hit->next_hsp ) { print "\t\tscore is ", $hsp->score, "\n"; } } } } } } # This example shows how to change a CGI parameter: $Bio::Tools::Run::RemoteBlast::HEADER{'MATRIX_NAME'} = 'BLOSUM45'; $Bio::Tools::Run::RemoteBlast::HEADER{'GAPCOSTS'} = '15 2'; # And this is how to delete a CGI parameter: delete $Bio::Tools::Run::RemoteBlast::HEADER{'FILTER'}; =head1 DESCRIPTION Class for remote execution of the NCBI Blast via HTTP. For a description of the many CGI parameters see: http://www.ncbi.nlm.nih.gov/BLAST/Doc/urlapi.html Various additional options and input formats are available. =head1 FEEDBACK =head2 Mailing Lists User feedback is an integral part of the evolution of this and other Bioperl modules. Send your comments and suggestions preferably to one of the Bioperl mailing lists. Your participation is much appreciated. bioperl-l@bioperl.org - General discussion http://bioperl.org/wiki/Mailing_lists - About the mailing lists =head2 Support Please direct usage questions or support issues to the mailing list: I rather than to the module maintainer directly. Many experienced and reponsive experts will be able look at the problem and quickly address it. Please include a thorough description of the problem with code and data examples if at all possible. =head2 Reporting Bugs Report bugs to the Bioperl bug tracking system to help us keep track the bugs and their resolution. Bug reports can be submitted via the web: http://bugzilla.bioperl.org =head1 AUTHOR Please do NOT contact Jason directly about this module. Please post to the bioperl mailing list (L). If you would like to be the official maintainer of this module, please volunteer on the list and we will make it official in this POD. First written by Jason Stajich, many others have helped keep it running. =head1 APPENDIX The rest of the documentation details each of the object methods. Internal methods are usually preceded with a _ =cut package Bio::Tools::Run::RemoteBlast; use vars qw($AUTOLOAD $URLBASE %HEADER %RETRIEVALHEADER $RIDLINE $MODVERSION %PUTPARAMS %GETPARAMS); use strict; use Bio::SeqIO; use IO::String; use Bio::SearchIO; use LWP; use HTTP::Request::Common; use base qw(Bio::Root::Root Bio::Root::IO); BEGIN { $MODVERSION = $Bio::Root::Version::VERSION; $URLBASE = 'http://blast.ncbi.nlm.nih.gov/Blast.cgi'; # In GET/PUTPARAMS the values are regexes which validate the input. %PUTPARAMS = ( 'AUTO_FORMAT' => '(Off|(Semi|Full)auto)', # Off, Semiauto, Fullauto 'COMPOSITION_BASED_STATISTICS' => '(0|1)', # yes, no on NCBI's site, but actually binary 0/1 'DATABASE' => '.*', 'DB_GENETIC_CODE' => '([1-9]|1[1-6]|2(1|2))', # 1..16,21,22 'DISPLAY_SORT' => '\d', 'ENDPOINTS' => '(yes|no)', # yes,no 'ENTREZ_QUERY' => '.*', 'EXPECT' => '\d+(\.\d+)?([eE]-\d+)?', # Positive double 'FILTER' => '[LRm]', # L or R or m 'GAPCOSTS' => '-?\d+(\.\d+)\s+i-?\d+(\.\d+)', # Two space separated float values 'GENETIC_CODE' => '([1-9]|1[1-6]|2(1|2))', # 1..16,21,22 'HITLIST_SIZE' => '\d+', # Positive integer 'I_THRESH' => '-?\d+(\.\d+)([eE]-\d+)?', # float 'LAYOUT' => '(One|Two)Windows?', # onewindow, twowindows 'LCASE_MASK' => '(yes|no)', # yes, no 'MATRIX_NAME' => '.*', 'NUCL_PENALTY' => '-\d+', # Negative integer 'NUCL_REWARD' => '-?\d+', # Integer 'OTHER_ADVANCED' => '.*', 'PERC_IDENT' => '\d\d+', # Integer, 0-99 inclusive 'PHI_PATTERN' => '.*', 'PROGRAM' => 't?blast[pnx]', # tblastp, tblastn, tblastx, blastp, blastn, blastx 'QUERY' => '.*', 'QUERY_FILE' => '.*', 'QUERY_BELIEVE_DEFLINE' => '(yes|no)', # yes, no 'QUERY_FROM' => '\d+', # Positive integer 'QUERY_TO' => '\d+', # Positive integer 'SEARCHSP_EFF' => '\d+', # Positive integer 'SERVICE' => '(plain|p[sh]i|(rps|mega)blast)', # plain,psi,phi,rpsblast,megablast 'SHORT_QUERY_ADJUST' => '(true|false)', 'THRESHOLD' => '-?\d+', # Integer 'UNGAPPED_ALIGNMENT' => '(yes|no)', # yes, no 'WORD_SIZE' => '\d+' # Positive integer ); %GETPARAMS = ( 'ALIGNMENTS' => '\d+', # Positive integer 'ALIGNMENT_VIEW' => '(Pairwise|(Flat)?QueryAnchored(NoIdentities)?|Tabular)', # Pairwise, QueryAnchored, QueryAnchoredNoIdentities, # FlatQueryAnchored, FlatQueryAnchoredNoIdentities, Tabular 'DATABASE_SORT' => '\d', 'DESCRIPTIONS' => '\d+', # Positive integer 'ENTREZ_LINKS_NEW_WINDOW' => '(yes|no)', # yes, no 'EXPECT_LOW' => '\d+(\.\d+)?([eE]-\d+)?', # Positive double 'EXPECT_HIGH' => '\d+(\.\d+)?([eE]-\d+)?', # Positive double 'FORMAT_ENTREZ_QUERY' => '', 'FORMAT_OBJECT' => '(Alignment|Neighbors|PSSM|SearchInfo|TaxBlast(Parent|MultiFrame)?)', # Alignment, Neighbors, PSSM, SearchInfo # TaxBlast, TaxblastParent, TaxBlastMultiFrame 'FORMAT_TYPE' => '((HT|X)ML|ASN\.1|Text)', # HTML, Text, ASN.1, XML 'NCBI_GI' => '(yes|no)', # yes, no 'NEW_VIEW' => '(true|false)', 'RID' => '.*', 'RESULTS_FILE' => '(yes|no)', # yes, no 'SERVICE' => '(plain|p[sh]i|(rps|mega)blast)', # plain,psi,phi,rpsblast,megablast 'SHOW_OVERVIEW' => '(yes|no)' # yes, no ); # Default values go in here for PUT %HEADER = ( 'CMD' => 'Put', 'FORMAT_OBJECT' => 'Alignment', 'COMPOSITION_BASED_STATISTICS' => 'off', 'DATABASE' => 'nr', 'EXPECT' => '1e-3', 'FILTER' => 'L', 'PROGRAM' => 'blastp', 'SERVICE' => 'plain' ); # Default values go in here for GET %RETRIEVALHEADER = ( 'CMD' => 'Get', 'ALIGNMENTS' => '50', 'ALIGNMENT_VIEW' => 'Pairwise', 'DESCRIPTIONS' => '100', 'FORMAT_TYPE' => 'Text', ); $RIDLINE = 'RID\s+=\s+(\S+)'; } sub new { my ($caller, @args) = @_; # chained new my $self = $caller->SUPER::new(@args); # so that tempfiles are cleaned up $self->_initialize_io(); my ($prog, $data, $readmethod, $url_base) = $self->_rearrange([qw(PROG DATA READMETHOD URL_BASE)], @args); # Use these two parameters for backward-compatibility. # Overridden by PROGRAM and DATABASE if supplied. $self->submit_parameter('PROGRAM',$prog) if $prog; $self->submit_parameter('DATABASE',$data) if $data; $readmethod = 'SearchIO' unless defined $readmethod; $self->readmethod($readmethod); # Now read the rest of the parameters and set them all # PUT parameters first my @putValues = $self->_rearrange([keys %PUTPARAMS],@args); my %putNames; @putNames{keys %PUTPARAMS} = @putValues; foreach my $putName (keys %putNames) { $self->submit_parameter($putName,$putNames{$putName}); } # GET parameters second my @getValues = $self->_rearrange([keys %GETPARAMS],@args); my %getNames; @getNames{keys %GETPARAMS} = @getValues; foreach my $getName (keys %getNames) { $self->retrieve_parameter($getName,$getNames{$getName}); } # private variable to keep track of total rids $self->{'_total_rids'} = 0; $url_base ||= $URLBASE; # default to regular NCBI BLAST URL $self->set_url_base($url_base); return $self; } =head2 retrieve_parameter Title : retrieve_parameter Usage : my $db = $self->retrieve_parameter Function: Get/Set the named parameter for the retrieve_blast operation. Returns : string Args : $name : name of GET parameter $val : optional value to set the parameter to =cut sub retrieve_parameter { my ($self, $name, $val) = @_; $name = uc($name); $self->throw($name." is not a valid GET parameter.") unless exists $GETPARAMS{$name}; if (defined $val) { my $regex = $GETPARAMS{$name}; $val =~ m/^$regex$/i or $self->throw("Value ".$val." for GET parameter ".$name." does not match expression ".$regex.". Rejecting."); $RETRIEVALHEADER{$name} = $val; } return $RETRIEVALHEADER{$name}; } =head2 submit_parameter Title : submit_parameter Usage : my $db = $self->submit_parameter Function: Get/Set the named parameter for the submit_blast operation. Returns : string Args : $name : name of PUT parameter $val : optional value to set the parameter to =cut sub submit_parameter { my ($self, $name, $val) = @_; $name = uc($name); $self->throw($name." is not a valid PUT parameter.") unless exists $PUTPARAMS{$name}; if (defined $val) { my $regex = $PUTPARAMS{$name}; $val =~ m/^$regex$/i or $self->throw("Value ".$val." for PUT parameter ".$name." does not match expression ".$regex.". Rejecting."); $HEADER{$name} = $val; } return $HEADER{$name}; } =head2 header Title : header Usage : my $header = $self->header Function: Get HTTP header for blast query Returns : string Args : none =cut sub header { my ($self) = @_; return %HEADER; } =head2 readmethod Title : readmethod Usage : my $readmethod = $self->readmethod Function: Get/Set the method to read the blast report Returns : string Args : string [ blast, blasttable, xml ] =cut sub readmethod { my ($self, $val) = @_; if( defined $val ) { if ($val =~ /bplite/i) { $self->throw("Use of Bio::Tools::BPlite is deprecated; use Bio::SearchIO modules instead"); } $self->{'_readmethod'} = $val; } return $self->{'_readmethod'}; } =head2 program Title : program Usage : my $prog = $self->program Function: Get/Set the program to run. Retained for backwards-compatibility. Returns : string Args : string [ blastp, blastn, blastx, tblastn, tblastx ] =cut sub program { my ($self, $val) = @_; return $self->submit_parameter('PROGRAM',$val); } =head2 database Title : database Usage : my $db = $self->database Function: Get/Set the database to search. Retained for backwards-compatibility. Returns : string Args : string [ swissprot, nr, nt, etc... ] =cut sub database { my ($self, $val) = @_; return $self->submit_parameter('DATABASE',$val); } =head2 expect Title : expect Usage : my $expect = $self->expect Function: Get/Set the E value cutoff. Retained for backwards-compatibility. Returns : string Args : string [ '1e-4' ] =cut sub expect { my ($self, $val) = @_; return $self->submit_parameter('EXPECT',$val); } =head2 ua Title : ua Usage : my $ua = $self->ua or $self->ua($ua) Function: Get/Set a LWP::UserAgent for use Returns : reference to LWP::UserAgent Object Args : none Comments: Will create a UserAgent if none has been requested before. =cut sub ua { my ($self, $value) = @_; if( ! defined $self->{'_ua'} ) { $self->{'_ua'} = LWP::UserAgent->new(env_proxy => 1, parse_head => 0); my $nm = ref($self); $nm =~ s/::/_/g; $self->{'_ua'}->agent("bioperl-$nm/$MODVERSION"); } return $self->{'_ua'}; } =head2 proxy Title : proxy Usage : $httpproxy = $db->proxy('http') or $db->proxy(['http','ftp'], 'http://myproxy' ) Function: Get/Set a proxy for use of proxy Returns : a string indicating the proxy Args : $protocol : an array ref of the protocol(s) to set/get $proxyurl : url of the proxy to use for the specified protocol =cut sub proxy { my ($self,$protocol,$proxy) = @_; return if ( !defined $self->ua || !defined $protocol || !defined $proxy ); return $self->ua->proxy($protocol,$proxy); } sub add_rid { my ($self, @vals) = @_; foreach ( @vals ) { $self->{'_rids'}->{$_} = $self->{'_total_rids'}; $self->{'_total_rids'}++; } return scalar keys %{$self->{'_rids'}}; } sub remove_rid { my ($self, @vals) = @_; foreach ( @vals ) { delete $self->{'_rids'}->{$_}; } return scalar keys %{$self->{'_rids'}}; } sub each_rid { my ($self) = @_; # sort on key value, a little tricky... my @sort_rids = sort {$self->{'_rids'}->{$a} <=> $self->{'_rids'}->{$b}} keys %{$self->{'_rids'}}; return @sort_rids; } =head2 submit_blast Title : submit_blast Usage : $self->submit_blast([$seq1,$seq2]); Function: Submit blast jobs to ncbi blast queue on sequence(s) Returns : Blast report object as defined by $self->readmethod Args : input can be: * sequence object * array ref of sequence objects * filename of file containing fasta formatted sequences =cut sub submit_blast { my ($self, $input) = @_; my @seqs = $self->_load_input($input); my $url_base = $self->get_url_base; return 0 unless ( @seqs ); my $tcount = 0; my %header = $self->header; $header{$_} ||= $RETRIEVALHEADER{$_} foreach (keys %RETRIEVALHEADER); foreach my $seq ( @seqs ) { #If query has a fasta header, the output has the query line. $header{'QUERY'} = ">".(defined $seq->display_id() ? $seq->display_id() : ""). " ".(defined $seq->desc() ? $seq->desc() : "")."\n".$seq->seq(); my $request = POST $url_base, [%header]; $self->warn($request->as_string) if ( $self->verbose > 0); my $response = $self->ua->request( $request); if( $response->is_success ) { my @subdata = split(/\n/, $response->content ); my $count = 0; foreach ( @subdata ) { if( /$RIDLINE/ ) { $count++; $self->debug("RID: $1\n"); $self->add_rid($1); last; } } if( $count == 0 ) { $self->warn("req was ". $request->as_string() . "\n"); $self->warn(join('', @subdata)); } $tcount += $count; } else { # should try and be a little more verbose here $self->warn("req was ". $request->as_string() . "\n" . $response->error_as_HTML); $tcount = -1; } } return $tcount; } =head2 retrieve_blast Title : retrieve_blast Usage : my $blastreport = $blastfactory->retrieve_blast($rid); Function: Attempts to retrieve a blast report from remote blast queue Returns : -1 on error, 0 on 'job not finished', Bio::SearchIO object Args : Remote Blast ID (RID) =cut sub retrieve_blast { my($self, $rid) = @_; my ($fh,$tempfile) = $self->tempfile(); close $fh; #explicit close my $url_base = $self->get_url_base; my %hdr = %RETRIEVALHEADER; $hdr{'RID'} = $rid; my $req = POST $url_base, [%hdr]; $self->debug("retrieve request is " . $req->as_string()); my $response = $self->ua->request($req, $tempfile); if( $response->is_success ) { if( $self->verbose > 0 ) { #print content of reply if verbose > 1 open(my $DEBUG, $tempfile) || $self->throw("cannot open $tempfile"); while(<$DEBUG>) { print $_; } close $DEBUG; } open(my $TMP, $tempfile) || $self->throw("Error opening $tempfile"); ## if proper reply my $waiting = 1; my $s = 0; my $got_content = 0; my $is_tabular = 0; while(my $line = <$TMP>) { if ($line =~ /./) { $got_content = 1; } if($line =~ /<\?xml version=/ ) { # xml time $waiting = 0; $self->readmethod('blastxml'); last; } if($line =~ /QBlastInfoBegin/i ) { $s = 1; } elsif( $s ) { if($line =~ /Status=(WAITING|ERROR|READY)/i ) { my $status = $1; if( $status eq 'WAITING' ) { $waiting = 1; } elsif( $status eq 'ERROR' ) { close($TMP); open(my $ERR, "<$tempfile") or $self->throw("cannot open file $tempfile"); $self->warn(join("", <$ERR>)); close $ERR; return -1; } elsif( $status eq 'READY' ) { $waiting = 0; last; } else { $self->warn("Unknown status $1:\n"); last; } } } elsif ($line =~ /ERROR/i ) { close($TMP); open(my $ERR, "<$tempfile") or $self->throw("cannot open file $tempfile"); $self->warn(join("", <$ERR>)); close $ERR; return -1; } } close($TMP); if( ! $waiting ) { my $blastobj; my $mthd = $self->readmethod; $mthd = ($mthd =~ /blasttable/i) ? 'blasttable' : ($mthd =~ /xml/i) ? 'blastxml' : ($mthd =~ /pull/i) ? 'blast_pull' : 'blast'; $blastobj = Bio::SearchIO->new( -file => $tempfile, -format => $mthd); ## store filename in object ## $self->file($tempfile); return $blastobj; } elsif (!$got_content) { # server returned no content, can't be good $self->warn("Server failed to return any data"); return -1 } else { # still working return 0; } } else { $self->warn($response->error_as_HTML); return -1; } } =head2 save_output Title : saveoutput Usage : my $saveoutput = $self->save_output($filename) Function: Method to save the blast report Returns : 1 (throws error otherwise) Args : string [rid, filename] =cut sub save_output { my ($self, $filename) = @_; if( ! defined $filename ) { $self->throw("Can't save blast output. You must specify a filename to save to."); } my $blastfile = $self->file; #open temp file and output file, have to filter out some HTML open(my $TMP, $blastfile) or $self->throw("cannot open $blastfile"); open(my $SAVEOUT, ">", $filename) or $self->throw("cannot open $filename"); my $seentop = 0; while(<$TMP>) { next if (/
/);
		if(/^(?:[T]?BLAST[NPX])\s*.+$/i ||
           /^RPS-BLAST\s*.+$/i ||
           /<\?xml\sversion=/ ||
           /^#\s+(?:[T]?BLAST[NPX])\s*.+$/) {
			$seentop=1;
		} 
        next if !$seentop;
		if( $seentop ) {
			print $SAVEOUT $_;
		}
	}
	return 1;
}

sub _load_input {
	my ($self, $input) = @_;

	if( ! defined $input ) {
		$self->throw("Calling remote blast with no input");
	}
	my @seqs;
	if( ! ref $input ) {
		if( -e $input ) {
			my $seqio = Bio::SeqIO->new(-format => 'fasta',
												-file => $input);
			while( my $seq = $seqio->next_seq ) {
				push @seqs, $seq;
			}
		} else {
			$self->throw("Input $input was not a valid filename");
		}
	} elsif( ref($input) =~ /ARRAY/i ) {
		foreach ( @$input ) {
			if( ref($_) && $_->isa('Bio::PrimarySeqI') ) {
				push @seqs, $_;
			} else {
				$self->warn("Trying to add a " . ref($_) .
								" but expected a Bio::PrimarySeqI");
			}
		}
		if( ! @seqs) {
			$self->throw("Did not pass in valid input -- no sequence objects found");
		}
	} elsif( $input->isa('Bio::PrimarySeqI') ) {
		push @seqs, $input;
	}
	return @seqs;
}

=head2 set_url_base

 Title   : set_url_base
 Usage   : $self->set_url_base($url)
 Function: Method to override the default NCBI BLAST database
 Returns : None
 Args    : string (database url like
 NOTE    : This is highly experimental; we cannot maintain support on
           databases other than the default NCBI database at this time

=cut

sub set_url_base {
    my $self = shift;
    $self->{'_urlbase'} = shift if @_;
}

=head2 get_url_base

 Title   : get_url_base
 Usage   : my $url = $self->set_url_base
 Function: Get the current URL for BLAST database searching
 Returns : string (URL used for remote blast searches)
 Args    : None

=cut

sub get_url_base {
    my $self = shift;
    return $self->{'_urlbase'};
}

1;

__END__