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# -*-Perl-*- mode (to keep my emacs happy)
# $Id: SeqIO.t,v 1.77 2003/12/17 13:14:28 heikki Exp $
use strict;
use vars qw($DEBUG $TESTCOUNT);
$DEBUG = $ENV{'BIOPERLDEBUG'} || 0;
BEGIN {
eval { require Test; };
if( $@ ) {
use lib 't';
}
use Test;
$TESTCOUNT = 235;
plan tests => $TESTCOUNT;
}
use Bio::Seq;
use Bio::SeqIO;
use Bio::SeqIO::MultiFile;
use Bio::Root::IO;
use Bio::Annotation::Collection;
ok(1);
# Set to -1 for release version, so warnings aren't printed
my $verbosity = $DEBUG ? 1 : -1;
#
# Basic read and/or write tests
#
sub read_write {
my $format = shift;
my $seq;
my $str = Bio::SeqIO->new(-file=> Bio::Root::IO->catfile("t","data","test.$format"),
'-format' => $format);
ok $seq = $str->next_seq();
print "Sequence 1 of 2 from $format stream:\n", $seq->seq, "\n\n" if $DEBUG;
unless ($format eq 'raw') {
ok $seq->id, 'roa1_drome',"ID for format $format";
ok $seq->length, 358;
}
unless ($format eq 'gcg') { # GCG file can contain only one sequence
ok $seq = $str->next_seq();
print "Sequence 2 of 2 from $format stream:\n", $seq->seq, $seq->seq, "\n" if $DEBUG;
}
my $out = Bio::SeqIO->new('-file'=> ">". Bio::Root::IO->catfile("t","data","$format.out"),
'-format' => $format);
ok $out->write_seq($seq);
if ($format eq 'fasta') {
my $id_type;
ok($id_type = $out->preferred_id_type('accession.version'), 'accession.version');
}
}
my @formats = qw(gcg fasta raw pir tab);
foreach my $format (@formats) {
print "======== $format ========\n" if $DEBUG;
read_write($format);
}
END {
map { unlink Bio::Root::IO->catfile("t","data","$_.out") } @formats
}
my ($str, $seq,$ast,$temp,$mf,$ent,$out); # predeclare variables for strict
# PIR testing
$str = Bio::SeqIO->new('-file' => Bio::Root::IO->catfile("t","data","seqfile.pir"),
'-format' => 'pir');
ok $str;
$out = new Bio::SeqIO(-format => 'pir', -fh => \*STDOUT);
while( $seq = $str->next_seq()) {
# ok($seq->id, qr /^[PF]1/ );
ok($seq->length > 1);
$out->write_seq($seq) if $verbosity > 0;
}
$out = undef;
$ast = Bio::SeqIO->new( '-format' => 'GenBank' ,
'-file' => Bio::Root::IO->catfile("t","data","roa1.genbank"));
$ast->verbose($verbosity);
my $as = $ast->next_seq();
ok $as->molecule, 'mRNA';
ok $as->alphabet, 'dna';
ok($as->primary_id, 3598416);
$ast = Bio::SeqIO->new( '-format' => 'genbank' ,
'-file' => Bio::Root::IO->catfile("t","data",
"NT_021877.gbk"));
$ast->verbose($verbosity);
$as = $ast->next_seq();
ok $as->molecule, 'DNA';
ok $as->alphabet, 'dna';
ok($as->primary_id, 37539616);
ok($as->accession_number, 'NT_021877');
my ($cds) = grep { $_->primary_tag eq 'CDS' } $as->get_SeqFeatures();
ok(($cds->get_tag_values('transl_except'))[1],
'(pos:complement(4224..4226),aa:OTHER)');
# embl
$ast = Bio::SeqIO->new( '-format' => 'embl' ,
'-file' => Bio::Root::IO->catfile("t","data","roa1.dat"));
$ast->verbose($verbosity);
$as = $ast->next_seq();
ok defined $as->seq;
ok($as->display_id, 'HSHNCPA1');
ok($as->accession_number, 'X79536');
ok($as->seq_version, 1);
ok($as->version, 1);
ok($as->desc, 'H.sapiens mRNA for hnRNPcore protein A1');
ok($as->molecule, 'RNA');
ok($as->alphabet, 'rna');
ok(scalar $as->all_SeqFeatures(), 4);
ok($as->length, 1198);
ok($as->species->binomial(), 'Homo sapiens');
$ent = Bio::SeqIO->new( '-file' => Bio::Root::IO->catfile("t","data","test.embl"),
'-format' => 'embl');
$seq = $ent->next_seq();
ok(defined $seq->seq(), 1,
'failure to read Embl with ^ location and badly split double quotes');
ok(scalar $seq->annotation->get_Annotations('reference'), 3);
$out = Bio::SeqIO->new('-file'=> ">embl.out",
'-format' => 'embl');
ok($out->write_seq($seq),1,
'failure to write Embl format with ^ < and > locations');
unlink("embl.out");
# kegg
my $kegg = Bio::SeqIO->new( '-format' => 'kegg' ,
'-file' => Bio::Root::IO->catfile("t","data","AHCYL1.kegg"));
ok($kegg);
$kegg = $kegg->next_seq();
ok($kegg);
ok($kegg->accession, '10768');
ok($kegg->display_id, 'AHCYL1');
ok($kegg->alphabet, 'dna');
ok($kegg->seq);
ok($kegg->translate->seq);
ok(($kegg->annotation->get_Annotations('description'))[0]->text,
'S-adenosylhomocysteine hydrolase-like 1 [EC:3.3.1.1]');
ok( (grep {$_->database eq 'KO'}
$kegg->annotation->get_Annotations('dblink'))[0]->comment,
'adenosylhomocysteinase' );
ok( (grep {$_->database eq 'PATH'}
$kegg->annotation->get_Annotations('dblink'))[0]->primary_id,
'hsa00271' );
# multifile
$mf = Bio::SeqIO::MultiFile->new( '-format' => 'Fasta' ,
'-files' =>
[ Bio::Root::IO->catfile("t","data","multi_1.fa"),
Bio::Root::IO->catfile("t","data","multi_2.fa")]);
ok defined $mf;
my $count = 0;
eval {
while( $seq = $mf->next_seq() ) {
$count++;
$temp = $seq->display_id;
}
};
ok( $count ,12);
$temp = undef;
# swissprot
$ast = Bio::SeqIO->new( '-verbosity' => $verbosity,
'-format' => 'swiss' ,
'-file' => Bio::Root::IO->catfile("t","data","roa1.swiss"));
$as = $ast->next_seq();
ok defined $as->seq;
ok($as->id, 'ROA1_HUMAN', "id is ".$as->id);
#ok($as->primary_id, 'ROA1');
skip($as->primary_id =~ /^Bio::Seq::/, $as->primary_id, 'ROA1');
ok($as->length, 371);
ok($as->alphabet, 'protein');
ok($as->division, 'HUMAN');
ok(scalar $as->all_SeqFeatures(), 16);
ok(scalar $as->annotation->get_Annotations('reference'), 11);
($ent, $seq, $out,$as) = undef;
#ace
{
my $t_file = Bio::Root::IO->catfile("t","data","test.ace");
my( $before );
{
local $/ = undef;
local *BEFORE;
open BEFORE, $t_file;
$before = <BEFORE>;
close BEFORE;
}
my $a_in = Bio::SeqIO->new( -FILE => $t_file, -FORMAT => 'ace');
my( @a_seq );
while (my $a = $a_in->next_seq) {
push(@a_seq, $a);
}
ok @a_seq, 3, 'wrong number of sequence objects';
my $esc_name = $a_seq[1]->display_id;
ok( $esc_name , 'Name; 4% strewn with \ various / escaped characters',
"bad unescaping of characters, $esc_name");
ok $a_seq[0]->alphabet, 'protein', 'alphabets incorrectly detected';
ok $a_seq[1]->alphabet, 'dna', 'alphabets incorrectly detected';
my $o_file = Bio::Root::IO->catfile("t","data","test.out.ace");
my $a_out = Bio::SeqIO->new( -FILE => "> $o_file", -FORMAT => 'ace');
my $a_out_ok = 1;
foreach my $a (@a_seq) {
$a_out->write_seq($a) or $a_out_ok = 0;
}
undef($a_out); # Flush to disk
ok $a_out_ok,1,'error writing sequence';
my( $after );
{
local $/ = undef;
local *AFTER;
open AFTER, $o_file;
$after = <AFTER>;
close AFTER;
}
unlink($o_file);
ok( ($before and $after and ($before eq $after)),1,
'test output file differs from input');
}
#
# streaming & Bio::RichSeq creation
#
my $stream = Bio::SeqIO->new('-file' => Bio::Root::IO->catfile("t","data","test.genbank"),
'-format' => 'GenBank');
$stream->verbose($verbosity);
my $seqnum = 0;
my $species;
my @cl;
my $lasts;
my @ids = qw(DDU63596 DDU63595 HUMBDNF);
my @tids = (44689, 44689, 9606);
my @tnames = ("Dictyostelium discoideum","Dictyostelium discoideum","Homo sapiens");
while($seq = $stream->next_seq()) {
if($seqnum < 3) {
ok $seq->display_id(), $ids[$seqnum];
$species = $seq->species();
@cl = $species->classification();
ok( $species->binomial(), $tnames[$seqnum],
'species parsing incorrect for genbank');
ok( $cl[3] ne $species->genus(), 1,
'genus duplicated in genbank parsing');
ok( $species->ncbi_taxid, $tids[$seqnum] );
}
$seqnum++;
$lasts = $seq;
}
ok $lasts->display_id(), "HUMBETGLOA";
my ($ref) = $lasts->annotation->get_Annotations('reference');
ok($ref->medline, 94173918);
$stream->close();
$stream = Bio::SeqIO->new(
'-file' => Bio::Root::IO->catfile("t","data","test.genbank.noseq"),
'-format' => 'GenBank',
);
$seqnum = 0;
while($seq = $stream->next_seq()) {
if($seqnum < 3) {
ok $seq->display_id(), $ids[$seqnum];
} elsif( $seq->display_id eq 'M37762') {
ok( ($seq->get_keywords())[0], 'neurotrophic factor');
}
$seqnum++;
}
ok $seqnum, 5, "Total number of sequences in test file";
$ent = Bio::SeqIO->new( '-file' => Bio::Root::IO->catfile("t","data","test.embl"),
'-format' => 'embl');
$ent->verbose($verbosity);
$seq = $ent->next_seq();
$species = $seq->species();
@cl = $species->classification();
ok( $cl[3] ne $species->genus(), 1, 'genus duplicated in EMBL parsing');
$ent->close();
$seq = Bio::SeqIO->new( '-format' => 'GenBank' ,
-file => Bio::Root::IO->catfile("t","data","testfuzzy.genbank"));
$seq->verbose($verbosity);
ok(defined($as = $seq->next_seq()));
my @features = $as->all_SeqFeatures();
ok(@features,21);
my $lastfeature = pop @features;
# this is a split location; the root doesn't have strand
ok($lastfeature->strand, undef);
my $location = $lastfeature->location;
$location->verbose(-1); # silence the warning of undef seq_id()
# see above; splitlocs roots do not have a strand really
ok($location->strand, undef);
ok($location->start, 83202);
ok($location->end, 84996);
my @sublocs = $location->sub_Location();
ok(@sublocs, 2);
my $loc = shift @sublocs;
ok($loc->start, 83202);
ok($loc->end, 83329);
ok($loc->strand, -1);
$loc = shift @sublocs;
ok($loc->start, 84248);
ok($loc->end, 84996);
ok($loc->strand,1);
$seq = Bio::SeqIO->new( '-format' => 'GenBank' ,
-file => ">".Bio::Root::IO->catfile("t","data","genbank.fuzzyout"));
$seq->verbose($verbosity);
ok($seq->write_seq($as));
unlink(Bio::Root::IO->catfile("t","data","genbank.fuzzyout"));
my $seqio = Bio::SeqIO->new( '-format' => 'swiss' ,
-file => Bio::Root::IO->catfile("t","data","swiss.dat"));
ok(defined( $seq = $seqio->next_seq));
# more tests to verify we are actually parsing correctly
skip($seq->primary_id =~ /^Bio::Seq/, $seq->primary_id, 'MA32');
ok($seq->display_id, 'MA32_HUMAN');
ok($seq->length, 282);
ok($seq->division, 'HUMAN');
ok($seq->alphabet, 'protein');
ok(scalar $seq->all_SeqFeatures(), 2);
my @genenames = qw(GC1QBP HABP1 SF2P32 C1QBP);
my ($ann) = $seq->annotation->get_Annotations('gene_name');
foreach my $gn ( $ann->get_all_values() ) {
ok ($gn, shift(@genenames));
}
ok $ann->value(-joins => [" AND "," OR "]), "GC1QBP OR HABP1 OR SF2P32 OR C1QBP";
# test for feature locations like ?..N
ok(defined( $seq = $seqio->next_seq));
skip($seq->primary_id =~ /^Bio::Seq/, $seq->primary_id, 'ACON');
ok($seq->display_id, 'ACON_CAEEL');
ok($seq->length, 788);
ok($seq->division, 'CAEEL');
ok($seq->alphabet, 'protein');
ok(scalar $seq->all_SeqFeatures(), 5);
foreach my $gn ( $seq->annotation->get_Annotations('gene_name') ) {
ok ($gn->value, 'F54H12.1');
}
# test species in swissprot -- this can be a n:n nightmare
ok ($seq = $seqio->next_seq());
my @sec_acc = $seq->get_secondary_accessions();
ok ($sec_acc[0], 'P29360');
ok ($sec_acc[1], 'Q63631');
ok ($seq->accession_number, 'P42655');
my @kw = $seq->get_keywords;
ok( $kw[0], 'Brain');
ok( $kw[1], 'Neurone');
ok ($kw[3], 'Multigene family');
ok ($seq->display_id, '143E_HUMAN');
ok ($seq->species->binomial, "Homo sapiens");
ok ($seq->species->common_name, "Human");
ok ($seq->species->ncbi_taxid, 9606);
ok ($seq = $seqio->next_seq());
ok ($seq->species->binomial, "Bos taurus");
ok ($seq->species->common_name, "Bovine");
ok ($seq->species->ncbi_taxid, 9913);
# multiple genes in swissprot
ok ($seq = $seqio->next_seq());
($ann) = $seq->annotation->get_Annotations("gene_name");
@genenames = qw(CALM1 CAM1 CALM CAM CALM2 CAM2 CAMB CALM3 CAM3 CAMC);
foreach my $gn ( $ann->get_all_values() ) {
ok ($gn, shift(@genenames));
}
ok $ann->value(-joins => [" AND "," OR "]),
"(CALM1 OR CAM1 OR CALM OR CAM) AND (CALM2 OR CAM2 OR CAMB) AND (CALM3 OR CAM3 OR CAMC)";
# test dos Linefeeds in gcg parser
$str = Bio::SeqIO->new('-file' => Bio::Root::IO->catfile("t","data","test_badlf.gcg"),
'-format' => 'GCG');
ok($str);
ok ( $seq = $str->next_seq());
ok(length($seq->seq) > 0 );
print "Sequence 1 of 1 from GCG stream:\n", $seq->seq, "\n" if( $DEBUG);
$str = new Bio::SeqIO(-format => 'genbank',
-file => Bio::Root::IO->catfile("t","data",
"AF165282.gb"),
-verbose => $verbosity);
$seq = $str->next_seq;
@features = $seq->all_SeqFeatures();
ok(@features, 5);
ok($features[0]->start, 1);
ok($features[0]->end, 226);
$location = $features[1]->location;
ok($location->isa('Bio::Location::SplitLocationI'));
@sublocs = $location->sub_Location();
ok(@sublocs, 29);
# version and primary ID - believe it or not, this wasn't working
ok ($seq->version, 1);
ok ($seq->seq_version, 1);
ok ($seq->primary_id, "5734104");
# test embl writing of a PrimarySeq
my $primaryseq = new Bio::PrimarySeq( -seq => 'AGAGAGAGATA',
-id => 'myid',
-desc => 'mydescr',
-alphabet => 'DNA',
-accession_number => 'myaccession');
my $embl = new Bio::SeqIO(-format => 'embl',
-verbose => $verbosity,
-file => ">primaryseq.embl");
ok($embl->write_seq($primaryseq));
my $scalar = "test";
eval {
$embl->write_seq($scalar);
};
ok ($@);
unlink("primaryseq.embl");
# revcomp split location
my $gb = new Bio::SeqIO(-format => 'genbank',
-file => Bio::Root::IO->catfile(qw(t data revcomp_mrna.gb)));
$seq = $gb->next_seq();
$gb = new Bio::SeqIO(-format => 'genbank',
-file => ">tmp_revcomp_mrna.gb");
$gb->write_seq($seq);
undef $gb;
ok(! -z "tmp_revcomp_mrna.gb");
# INSERT DIFFING CODE HERE
unlink("tmp_revcomp_mrna.gb");
# test secondary accessions in EMBL (bug #1332)
$seqio = new Bio::SeqIO(-format =>'embl', -file => Bio::Root::IO->catfile
( qw(t data ECAPAH02.embl)));
$seq = $seqio->next_seq;
ok($seq->accession_number, 'D10483');
ok($seq->seq_version, 2);
my @accs = $seq->get_secondary_accessions();
ok($accs[0], 'J01597');
ok($accs[-1], 'X56742');
$seqio = new Bio::SeqIO(-format => 'genbank',
-file => Bio::Root::IO->catfile(qw(t data
D10483.gbk)));
$seq = $seqio->next_seq;
@kw = $seq->get_keywords;
ok(scalar @kw, 118);
ok($kw[-1], 'yabO');
@sec_acc = $seq->get_secondary_accessions();
ok(scalar @sec_acc,14);
ok($sec_acc[-1], 'X56742');
### TPA TESTS - Thanks to Richard Adams ###
### test Third Party Annotation entries in EMBL/Gb format
# to ensure compatability with parsers.
###embl####
$str = new Bio::SeqIO(-format =>'embl', -file => Bio::Root::IO->catfile
( qw(t data BN000066-tpa.embl)));
$seq = $str->next_seq;
ok(defined $seq);
ok($seq->accession_number, 'BN000066');
ok($seq->alphabet, 'dna');
ok($seq->display_id, 'AGA000066');
ok($seq->length, 5195);
ok($seq->division, 'INV');
ok($seq->get_dates, 2);
ok($seq->keywords, 'acetylcholinesterase; achE1 gene; Third Party Annotation; TPA.');
ok($seq->seq_version, 1);
ok($seq->feature_count, 15);
my $spec_obj = $seq->species;
ok ($spec_obj->common_name, 'African malaria mosquito');
ok ($spec_obj->species, 'gambiae');
ok ($spec_obj->genus, 'Anopheles');
ok ($spec_obj->binomial, 'Anopheles gambiae');
my $ac = $seq->annotation;
my $reference = ($ac->get_Annotations('reference') )[1];
ok ($reference->title,'"A novel acetylcholinesterase gene in mosquitoes codes for the insecticide target and is non-homologous to the ace gene in Drosophila"');
ok ($reference->authors,'Weill M., Fort P., Berthomi eu A., Dubois M.P., Pasteur N., Raymond M.');
my $cmmnt = ($ac->get_Annotations('comment') )[0];
ok($cmmnt->text, 'see also AJ488492 for achE-1 from Kisumu strain Third Party Annotation Database: This TPA record uses Anopheles gambiae trace archive data (http://trace.ensembl.org) ');
## now genbank ##
$str = new Bio::SeqIO(-format =>'genbank', -file => Bio::Root::IO->catfile
( qw(t data BK000016-tpa.gbk)));
$seq = $str->next_seq;
ok(defined $seq);
ok(defined $seq->seq);
ok($seq->accession_number, 'BK000016');
ok($seq->alphabet, 'dna');
ok($seq->display_id, 'BK000016');
ok($seq->length, 1162);
ok($seq->division, 'ROD');
ok($seq->get_dates, 1);
ok($seq->keywords, 'Third Party Annotation; TPA');
ok($seq->desc, 'TPA: Mus musculus pantothenate kinase 4 mRNA, partial cds.');
ok($seq->seq_version, 1);
ok($seq->feature_count, 2);
$spec_obj = $seq->species;
ok ($spec_obj->common_name, 'Mus musculus (house mouse)');
ok ($spec_obj->species, 'musculus');
ok ($spec_obj->genus, 'Mus');
ok ($spec_obj->binomial, 'Mus musculus');
$ac = $seq->annotation;
$reference = ($ac->get_Annotations('reference') )[0];
ok ($reference->pubmed, '11479594');
ok ($reference->medline, '21372465');
# validate that what is written is what is read
my $testfile = "testtpa.gbk";
$out = new Bio::SeqIO(-file => ">$testfile",
-format => 'genbank');
$out->write_seq($seq);
$out->close();
$str = new Bio::SeqIO(-format =>'genbank',
-file => $testfile);
$seq = $str->next_seq;
ok(defined $seq);
ok(defined $seq->seq);
ok($seq->accession_number, 'BK000016');
ok($seq->alphabet, 'dna');
ok($seq->display_id, 'BK000016');
ok($seq->length, 1162);
ok($seq->division, 'ROD');
ok($seq->get_dates, 1);
ok($seq->keywords, 'Third Party Annotation; TPA');
ok($seq->desc, 'TPA: Mus musculus pantothenate kinase 4 mRNA, partial cds.');
ok($seq->seq_version, 1);
ok($seq->feature_count, 2);
$spec_obj = $seq->species;
ok ($spec_obj->common_name, 'Mus musculus (house mouse)');
ok ($spec_obj->species, 'musculus');
ok ($spec_obj->genus, 'Mus');
ok ($spec_obj->binomial, 'Mus musculus');
$ac = $seq->annotation;
$reference = ($ac->get_Annotations('reference') )[0];
ok ($reference->pubmed, '11479594');
ok ($reference->medline, '21372465');
unlink($testfile);
# bug #1487
$str = new Bio::SeqIO(-verbose => $verbosity,
-file => Bio::Root::IO->catfile
(qw(t data D12555.gbk)));
eval {
$seq = $str->next_seq;
};
ok(! $@ );
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