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# -*-Perl-*- Test Harness script for Bioperl
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# $Id: Analysis.t 15112 2008-12-08 18:12:38Z sendu $
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test_begin(-tests => 177);
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use_ok('Bio::Restriction::Enzyme');
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use_ok('Bio::Restriction::Enzyme::MultiCut');
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use_ok('Bio::Restriction::Enzyme::MultiSite');
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use_ok('Bio::Restriction::EnzymeCollection');
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use_ok('Bio::Restriction::Analysis');
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# Bio::Restriction::Enzyme
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my ($re, $seq, $iso, %meth, $microbe, $source, @vendors, @refs, $name);
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ok $re=Bio::Restriction::Enzyme->new(-enzyme=>'EcoRI', -site=>'G^AATTC');
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isa_ok($re, 'Bio::Restriction::EnzymeI');
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is $re->complementary_cut, 6;
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is $re->complementary_cut,5;
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is $re->site,'G^AATTC';
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isa_ok($seq, 'Bio::PrimarySeqI');
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is $seq->seq, 'GAATTC';
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is $re->string,'GAATTC';
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is $re->revcom, 'GAATTC';
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is $re->recognition_length, 6;
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is $re->palindromic, 1;
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is $re->overhang, "5'";
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is $re->overhang_seq, 'AATT';
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is $re->is_ambiguous, 0;
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ok $re->compatible_ends($re);
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ok $re->isoschizomers('BamHI', 'AvaI'); # not really true :)
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is my @isos=$re->isoschizomers, 2;
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ok $re->purge_isoschizomers;
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is scalar($re->isoschizomers), 0;
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ok $re->methylation_sites(2,5); # not really true :)
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ok %meth = $re->methylation_sites;
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ok $re->purge_methylation_sites;
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is scalar($re->methylation_sites), 0;
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ok $re->microbe('E. coli');
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ok $microbe = $re->microbe;
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is $microbe, "E. coli";
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ok $re->source("Rob"); # not really true :)
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ok $source = $re->source;
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ok $re->vendors('NEB'); # my favorite
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ok @vendors = $re->vendors;
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is $vendors[0], "NEB";
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is scalar($re->vendors), 0;
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ok $re->references('Rob et al');
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ok @refs = $re->references;
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is $refs[0], "Rob et al";
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$re->purge_references;
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is scalar($re->references), 0;
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ok $re->name('BamHI');
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eval {$re->is_prototype};
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like($@, qr/Can't unequivocally assign prototype based on input format alone/, 'bug 2179');
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is $re->is_prototype(0), 0;
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is $re->is_prototype, 0;
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is $re->is_prototype(1), 1;
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is $re->is_prototype, 1;
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is $re->prototype_name, $re->name;
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ok ! $re->is_prototype(0);
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is $re->prototype_name('XxxI'), 'XxxI';
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is $re->prototype_name, 'XxxI';
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ok $re->seq->seq('RCATGY');
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ok my $re2 = $re->clone;
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is $re->name, $re2->name;
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ok $re = Bio::Restriction::Enzyme->new(-enzyme=>'AciI',
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is $re->palindromic, 0;
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is $re->is_palindromic, 0;
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# Bio::Restriction::Enzyme::MultiSite
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ok $re=Bio::Restriction::Enzyme::MultiSite->new(-enzyme=>'TaqII',
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-complementary_cut=>15
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ok $re2=Bio::Restriction::Enzyme::MultiSite->new(-enzyme=>'TaqII',
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-complementary_cut=>15
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isa_ok( $re, 'Bio::Restriction::EnzymeI');
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isa_ok( $re2, 'Bio::Restriction::EnzymeI');
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ok $re->others($re2);
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ok $re2->others($re);
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ok my $re3 = $re->clone;
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is $re->name , $re3->name; # wouldn't this be a circular reference???
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#print Dumper $re, $re3;exit;
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# Bio::Restriction::Enzyme::MultiCut
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#Hin4I has four cut sites [(8/13)GAYNNNNNVTC(13/8)],
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ok $re = Bio::Restriction::Enzyme::MultiCut->new(-enzyme=>'Hin4I',
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-site=>'GAYNNNNNVTC',
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-complementary_cut=>-13
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ok $re2 = Bio::Restriction::Enzyme::MultiCut->new(-enzyme=>'Hin4I',
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-site=>'GAYNNNNNVTC',
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-complementary_cut=>8
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isa_ok($re, 'Bio::Restriction::EnzymeI');
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isa_ok($re2, 'Bio::Restriction::EnzymeI');
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ok $re->others($re2);
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ok $re2->others($re);
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ok $re3 = $re->clone;
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is $re->name, $re3->name;
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#print Dumper $re, $re3;exit;
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# Bio::Restriction::EnzymeCollection
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my ($collection, $enz, $new_set);
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ok $collection = Bio::Restriction::EnzymeCollection->new(-empty=>1);
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is $collection->each_enzyme, 0;
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$collection = Bio::Restriction::EnzymeCollection->new;
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is $collection->each_enzyme, 532;
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is $collection->each_enzyme, 532;
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ok $enz = $collection->get_enzyme('AclI');
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isa_ok($enz, 'Bio::Restriction::Enzyme');
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is my @enzymes=$collection->available_list, 532;
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ok $new_set = $collection->blunt_enzymes;
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isa_ok($enz, 'Bio::Restriction::Enzyme');
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is $new_set->each_enzyme, 114;
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#map {print $_->name, ": ", $_->cutter, "\n"; } $collection->each_enzyme;
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ok $new_set = $collection->cutters(8);
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is $new_set->each_enzyme, 17;
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ok $new_set=$collection->cutters(-start => 8, -end => 8);
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is $new_set->each_enzyme, 17;
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ok $new_set=$collection->cutters(-start => 6, -end => 8);
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is $new_set->each_enzyme, 293;
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ok $new_set=$collection->cutters(-start => 6, -end => 8, -exclusive => 1);
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is $new_set->each_enzyme, 10;
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ok $new_set = $collection->cutters([4,8]);
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is $new_set->each_enzyme, 129;
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# bug 2128; enhancement request to pass array ref of sizes
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# Restriction::Analysis
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ok my $seqio=Bio::SeqIO->new(-file=>test_input_file('dna1.fa'),
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ok $seq=$seqio->next_seq;
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ok my $ra = Bio::Restriction::Analysis->new(-seq=>$seq);
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ok my $uniq = $ra->unique_cutters;
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is $ra->unique_cutters->each_enzyme, 42, 'number of unique cutters';
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is $ra->fragments('RsaI'), 2, 'number of RsaI fragments';
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is $ra->max_cuts, 9, 'number of maximum cutters';
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is $ra->zero_cutters->each_enzyme, 477, 'number of zero cutters';
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is $ra->cutters->each_enzyme, 55, 'number of cutters';
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is $ra->cutters(3)->each_enzyme, 8, 'number of 3x cutters';
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is $ra->fragments('MseI'), 4, '4 MseI fragments';
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is $ra->cuts_by_enzyme('MseI'), 3, '3 MseI cut sites';
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#my $z = $ra->cutters(3);
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#my $out=Bio::Restriction::IO->new;
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is $ra->fragments('PspEI'), 2, 'expected 2 PspEI fragments';
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is $ra->cuts_by_enzyme('PspEI'), 1;
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is $ra->cuts_by_enzyme('XxxI'), undef;
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is my @ss = $ra->sizes('PspEI'), 2, 'expected 2 sizes for PspEI';
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is $ss[0] + $ss[1], $seq->length;
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is $ra->fragments('MwoI'), 1, 'not circular expected 1 fragments for MwoI as it doesnt cut';
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# circularise the sequence, regenerate the cuts and test again
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# note that there is one less fragment now!
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ok $seq->is_circular(1);
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# we need to regenerate all the cuts
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is $ra->fragments('RsaI'), 1, 'number of RsaI fragments';
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is $ra->fragments('MseI'), 3, '3 circular MseI fragments';
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is $ra->cuts_by_enzyme('MseI'), 3, '3 circular MseI cut sites';
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is $ra->fragments('AciI'), 1, 'number for AciI a non-palindromic enzyme';
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is $ra->fragments('MwoI'), 1, '1 fragment for MwoI as it cuts across the circ point';
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ok my @rb=($collection->get_enzyme("AluI"), $collection->get_enzyme("MseI"), $collection->get_enzyme("MaeIII"));
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# test multiple digests
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ok my $rbc=Bio::Restriction::EnzymeCollection->new(-empty=>1);
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ok $rbc->enzymes(@rb);
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ok $ra->cut('multiple', $rbc);
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is $ra->fragments('multiple_digest'),7, '7 fragments in the multiple digest';
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is my @pos=$ra->positions('multiple_digest'),7, '7 positions in the multiple digest';
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is my @ssm = $ra->sizes('multiple_digest'),7, '7 sizes in the multiple digest';
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map {$check_len+=$_}@ssm;
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is $check_len, $seq->length;
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# now test the non-palindromic part
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# HindI is a non palindromic enzyme that cuts 9 times
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is $ra->positions('HindI'), 9, ' expected 9 cuts for HindI';
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# now we need to test the fragment maps
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# lets do this for HindI
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is my @fm=$ra->fragment_maps('HindI'), 9, 'expect 9 fragment maps for HindI';
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foreach my $fm (@fm) {
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is exists $fm->{'seq'}, 1, "sequence for ".$fm->{'seq'};
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is exists $fm->{'start'}, 1, "start at ".$fm->{'start'};
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is exists $fm->{'end'}, 1, "end at ".$fm->{'end'};
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eval {$re = Bio::Restriction::Enzyme->new(
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-site => 'G^AATTE' );};
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like($@, qr(Unrecognized characters in site), 'bug 2139');