| BioPerl documentation | Contained in the BioPerl distribution. |
Bio::Search::Tiling::MapTiling - An implementation of an HSP tiling algorithm, with methods to obtain frequently-requested statistics
# get a BLAST $hit from somewhere, then
$tiling = Bio::Search::Tiling::MapTiling->new($hit);
# stats
$numID = $tiling->identities();
$numCons = $tiling->conserved();
$query_length = $tiling->length('query');
$subject_length = $tiling->length('subject'); # or...
$subject_length = $tiling->length('hit');
# get a visual on the coverage map
print $tiling->coverage_map_as_text('query',$context,'LEGEND');
# tilings
$context = $tiling->_context( -type => 'subject', -strand=> 1, -frame=>1);
@covering_hsps_for_subject = $tiling->next_tiling('subject',$context);
$context = $tiling->_context( -type => 'query', -strand=> -1, -frame=>0);
@covering_hsps_for_query = $tiling->next_tiling('query', $context);
Frequently, users want to use a set of high-scoring pairs (HSPs) obtained from a BLAST or other search to assess the overall level of identity, conservation, or coverage represented by matches between a subject and a query sequence. Because a set of HSPs frequently describes multiple overlapping sequence fragments, a simple summation of statistics over the HSPs will generally overestimate those statistics. To obtain an accurate estimate of global hit statistics, a 'tiling' of HSPs onto either the subject or the query sequence must be performed, in order to properly correct for this.
This module will execute a tiling algorithm on a given hit based on an interval decomposition I'm calling the "coverage map". Internal object methods compute the various statistics, which are then stored in appropriately-named public object attributes. See Bio::Search::Tiling::MapTileUtils for more info on the algorithm.
In BLASTX, TBLASTN, and TBLASTX reports, strand and frame information are reported for the query, subject, or query and subject, respectively, for each HSP. Tilings for these sequence types are only meaningful when they include HSPs in the same strand and frame, or "context". So, in these situations, the context must be specified in the method calls or the methods will throw.
Contexts are specified as strings: [ 'all' | [m|p][_|0|1|2] ], where
all = all HSPs (will throw if context must be specified), m = minus
strand, p = plus strand, and _ = no frame info, 0,1,2 = respective
(absolute) frame. The _make_context_key method will convert a (strand,
frame) specification to a context string, e.g.:
$context = $self->_context(-type=>'query', -strand=>-1, -frame=>-2);
returns m2.
The contexts present among the HSPs in a hit are identified and stored
for convenience upon object construction. These are accessed off the
object with the contexts method. If contexts don't apply for the
given report, this returns ('all').
The experimental method get_tiled_alns in ALIGNMENTS will use a tiling to concatenate tiled hsps into a series of Bio::SimpleAlign objects:
@alns = $tiling->get_tiled_alns($type, $context);
Each alignment contains two sequences with ids 'query' and 'subject',
and consists of a concatenation of tiling HSPs which overlap or are
directly adjacent. The alignment are returned in $type sequence
order. When HSPs overlap, the alignment sequence is taken from the HSP
which comes first in the coverage map array.
The sequences in each alignment contain features (even though they are Bio::LocatableSeq objects) which map the original query/subject coordinates to the new alignment sequence coordinates. You can determine the original BLAST fragments this way:
$aln = ($tiling->get_tiled_alns)[0];
$qseq = $aln->get_seq_by_id('query');
$hseq = $aln->get_seq_by_id('subject');
foreach my $feat ($qseq->get_SeqFeatures) {
$org_start = ($feat->get_tag_values('query_start'))[0];
$org_end = ($feat->get_tag_values('query_end'))[0];
# original fragment as represented in the tiled alignment:
$org_fragment = $feat->seq;
}
foreach my $feat ($hseq->get_SeqFeatures) {
$org_start = ($feat->get_tag_values('subject_start'))[0];
$org_end = ($feat->get_tag_values('subject_end'))[0];
# original fragment as represented in the tiled alignment:
$org_fragment = $feat->seq;
}
The major calculations are made just-in-time, and then memoized. So, for example, for a given MapTiling object, a coverage map would usually be calculated only once (for the query), and at most twice (if the subject perspective is also desired), and then only when a statistic is first accessed. Afterward, the map and/or any statistic is read from storage. So feel free to call the statistic methods frequently if it suits you.
User feedback is an integral part of the evolution of this and other Bioperl modules. Send your comments and suggestions preferably to the Bioperl mailing list. Your participation is much appreciated.
bioperl-l@bioperl.org - General discussion http://bioperl.org/wiki/Mailing_lists - About the mailing lists
Please direct usage questions or support issues to the mailing list:
bioperl-l@bioperl.org
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.
Report bugs to the Bioperl bug tracking system to help us keep track of the bugs and their resolution. Bug reports can be submitted via the web:
https://redmine.open-bio.org/projects/bioperl/
Email maj -at- fortinbras -dot- us
The rest of the documentation details each of the object methods. Internal methods are usually preceded with a _
Title : new
Usage : my $obj = new Bio::Search::Tiling::GenericTiling();
Function: Builds a new Bio::Search::Tiling::GenericTiling object
Returns : an instance of Bio::Search::Tiling::GenericTiling
Args : -hit => $a_Bio_Search_Hit_HitI_object
general filter function:
-hsp_filter => sub { my $this_hsp = shift;
...;
return 1 if $wanted;
return 0; }
Title : next_tiling
Usage : @hsps = $self->next_tiling($type);
Function: Obtain a tiling: a minimal set of HSPs covering the $type
('hit', 'subject', 'query') sequence
Example :
Returns : an array of HSPI objects
Args : scalar $type: one of 'hit', 'subject', 'query', with
'subject' an alias for 'hit'
Title : rewind_tilings
Usage : $self->rewind_tilings($type)
Function: Reset the next_tilings($type) iterator
Example :
Returns : True on success
Args : scalar $type: one of 'hit', 'subject', 'query';
default is 'query'
Title : get_tiled_alns
Usage : @alns = $tiling->get_tiled_alns($type, $context)
Function: Use a tiling to construct a minimal set of alignment
objects covering the region specified by $type/$context
by splicing adjacent HSP tiles
Returns : an array of Bio::SimpleAlign objects; see Note below
Args : scalar $type: one of 'hit', 'subject', 'query'
default is 'query'
scalar $context: strand/frame context string
Following $type and $context, an array of
ordered, tiled HSP objects can be specified; this is
the tiling that will directly the alignment construction
default -- the first tiling provided by a tiling iterator
Notes : Each returned alignment is a concatenation of adjacent tiles.
The set of alignments will cover all regions described by the
$type/$context pair in the hit. The pair of sequences in each
alignment have ids 'query' and 'subject', and each sequence
possesses SeqFeatures that map the original query or subject
coordinates to the sequence coordinates in the tiled alignment.
Title : identities
Usage : $tiling->identities($type, $action, $context)
Function: Retrieve the calculated number of identities for the invocant
Example :
Returns : value of identities (a scalar)
Args : scalar $type: one of 'hit', 'subject', 'query'
default is 'query'
option scalar $action: one of 'exact', 'est', 'fast', 'max'
default is 'exact'
option scalar $context: strand/frame context string
Note : getter only
Title : conserved
Usage : $tiling->conserved($type, $action)
Function: Retrieve the calculated number of conserved sites for the invocant
Example :
Returns : value of conserved (a scalar)
Args : scalar $type: one of 'hit', 'subject', 'query'
default is 'query'
option scalar $action: one of 'exact', 'est', 'fast', 'max'
default is 'exact'
option scalar $context: strand/frame context string
Note : getter only
Title : length
Usage : $tiling->length($type, $action, $context)
Function: Retrieve the total length of aligned residues for
the seq $type
Example :
Returns : value of length (a scalar)
Args : scalar $type: one of 'hit', 'subject', 'query'
default is 'query'
option scalar $action: one of 'exact', 'est', 'fast', 'max'
default is 'exact'
option scalar $context: strand/frame context string
Note : getter only
Title : frac
Usage : $tiling->frac($type, $denom, $action, $context, $method)
Function: Return the fraction of sequence length consisting
of desired kinds of pairs (given by $method),
with respect to $denom
Returns : scalar float
Args : -type => one of 'hit', 'subject', 'query'
-denom => one of 'total', 'aligned'
-action => one of 'exact', 'est', 'fast', 'max'
-context => strand/frame context string
-method => one of 'identical', 'conserved'
Note : $denom == 'aligned', return desired_stat/num_aligned
$denom == 'total', return desired_stat/_reported_length
(i.e., length of the original input sequences)
Note : In keeping with the spirit of Bio::Search::HSP::HSPI,
reported lengths of translated dna are reduced by
a factor of 3, to provide fractions relative to
amino acid coordinates.
Title : frac_identical
Usage : $tiling->frac_identical($type, $denom, $action, $context)
Function: Return the fraction of sequence length consisting
of identical pairs, with respect to $denom
Returns : scalar float
Args : -type => one of 'hit', 'subject', 'query'
-denom => one of 'total', 'aligned'
-action => one of 'exact', 'est', 'fast', 'max'
-context => strand/frame context string
Note : $denom == 'aligned', return conserved/num_aligned
$denom == 'total', return conserved/_reported_length
(i.e., length of the original input sequences)
Note : In keeping with the spirit of Bio::Search::HSP::HSPI,
reported lengths of translated dna are reduced by
a factor of 3, to provide fractions relative to
amino acid coordinates.
Note : This an alias that calls frac()
Title : frac_conserved
Usage : $tiling->frac_conserved($type, $denom, $action, $context)
Function: Return the fraction of sequence length consisting
of conserved pairs, with respect to $denom
Returns : scalar float
Args : -type => one of 'hit', 'subject', 'query'
-denom => one of 'total', 'aligned'
-action => one of 'exact', 'est', 'fast', 'max'
-context => strand/frame context string
Note : $denom == 'aligned', return conserved/num_aligned
$denom == 'total', return conserved/_reported_length
(i.e., length of the original input sequences)
Note : In keeping with the spirit of Bio::Search::HSP::HSPI,
reported lengths of translated dna are reduced by
a factor of 3, to provide fractions relative to
amino acid coordinates.
Note : This an alias that calls frac()
Title : frac_aligned
Aliases : frac_aligned_query - frac_aligned(-type=>'query',...)
frac_aligned_hit - frac_aligned(-type=>'hit',...)
Usage : $tiling->frac_aligned(-type=>$type,
-action=>$action,
-context=>$context)
Function: Return the fraction of input sequence length
that was aligned by the algorithm
Returns : scalar float
Args : -type => one of 'hit', 'subject', 'query'
-action => one of 'exact', 'est', 'fast', 'max'
-context => strand/frame context string
Title : num_aligned
Usage : $tiling->num_aligned(-type=>$type)
Function: Return the number of residues of sequence $type
that were aligned by the algorithm
Returns : scalar int
Args : -type => one of 'hit', 'subject', 'query'
-action => one of 'exact', 'est', 'fast', 'max'
-context => strand/frame context string
Note : Since this is calculated from reported coordinates,
not symbol string counts, it is already in terms of
"logical length"
Note : Aliases length()
Title : num_unaligned
Usage : $tiling->num_unaligned(-type=>$type)
Function: Return the number of residues of sequence $type
that were left unaligned by the algorithm
Returns : scalar int
Args : -type => one of 'hit', 'subject', 'query'
-action => one of 'exact', 'est', 'fast', 'max'
-context => strand/frame context string
Note : Since this is calculated from reported coordinates,
not symbol string counts, it is already in terms of
"logical length"
Title : range
Usage : $tiling->range(-type=>$type)
Function: Returns the extent of the longest tiling
as ($min_coord, $max_coord)
Returns : array of two scalar integers
Args : -type => one of 'hit', 'subject', 'query'
-context => strand/frame context string
Title : coverage_map
Usage : $map = $tiling->coverage_map($type)
Function: Property to contain the coverage map calculated
by _calc_coverage_map() - see that for
details
Example :
Returns : value of coverage_map_$type as an array
Args : scalar $type: one of 'hit', 'subject', 'query'
default is 'query'
Note : getter
Title : coverage_map_as_text
Usage : $tiling->coverage_map_as_text($type, $legend_flag)
Function: Format a text-graphic representation of the
coverage map
Returns : an array of scalar strings, suitable for printing
Args : $type: one of 'query', 'hit', 'subject'
$context: strand/frame context string
$legend_flag: boolean; add a legend indicating
the actual interval coordinates for each component
interval and hsp (in the $type sequence context)
Example : print $tiling->coverage_map_as_text('query',1);
Title : hit Usage : $tiling->hit Function: Example : Returns : The HitI object associated with the invocant Args : none Note : getter only
Title : hsps Usage : $tiling->hsps() Function: Container for the HSP objects associated with invocant Example : Returns : an array of hsps associated with the hit Args : on set, new value (an arrayref or undef, optional)
Title : contexts
Usage : @contexts = $tiling->context($type) or
@indices = $tiling->context($type, $context)
Function: Retrieve the set of available contexts in the hit,
or the indices of hsps having the given context
(integer indices for the array returned by $self->hsps)
Returns : array of scalar context strings or
array of scalar positive integers
undef if no hsps in given context
Args : $type: one of 'query', 'hit', 'subject'
optional $context: context string
Title : mapping
Usage : $tiling->mapping($type)
Function: Retrieve the mapping coefficient for the sequence type
based on the underlying algorithm
Returns : scalar integer (mapping coefficient)
Args : $type: one of 'query', 'hit', 'subject'
Note : getter only (set in constructor)
Title : default_context
Usage : $tiling->default_context($type)
Function: Retrieve the default strand/frame context string
for the sequence type based on the underlying algorithm
Returns : scalar string (context string)
Args : $type: one of 'query', 'hit', 'subject'
Note : getter only (set in constructor)
Title : algorithm
Usage : $tiling->algorithm
Function: Retrieve the algorithm name associated with the
invocant's hit object
Returns : scalar string
Args : none
Note : getter only (set in constructor)
See Bio::Search::Tiling::MapTileUtils for lower level calculation methods.
Title : _calc_coverage_map
Usage : $tiling->_calc_coverage_map($type)
Function: Calculates the coverage map for the object's associated
hit from the perspective of the desired $type (see Args:)
and sets the coverage_map() property
Returns : True on success
Args : optional scalar $type: one of 'hit'|'subject'|'query'
default is 'query'
Note : The "coverage map" is an array with the following format:
( [ $component_interval => [ @containing_hsps ] ], ... ),
where $component_interval is a closed interval (see
DESCRIPTION) of the form [$a0, $a1] with $a0 <= $a1, and
@containing_hsps is an array of all HspI objects in the hit
which completely contain the $component_interval.
The set of $component_interval's is a disjoint decomposition
of the minimum set of minimal intervals that completely
cover the hit's HSPs (from the perspective of the $type)
Note : This calculates the map for all strand/frame contexts available
in the hit
Title : _calc_stats
Usage : $tiling->_calc_stats($type, $action, $context)
Function: Calculates [estimated] tiling statistics (identities, conserved sites
length) and sets the public accessors
Returns : True on success
Args : scalar $type: one of 'hit', 'subject', 'query'
default is 'query'
optional scalar $action: requests calculation method
currently one of 'exact', 'est', 'fast', 'max'
option scalar $context: strand/frame context string
Note : Action: The statistics are calculated by summing quantities
over the disjoint component intervals, taking into account
coverage of those intervals by multiple HSPs. The action
tells the algorithm how to obtain those quantities--
'exact' will use Bio::Search::HSP::HSPI::matches
to count the appropriate segment of the homology string;
'est' will estimate the statistics by multiplying the
fraction of the HSP overlapped by the component interval
(see MapTileUtils) by the BLAST-reported identities/postives
(this may be convenient for BLAST summary report formats)
* Both exact and est take the average over the number of HSPs
that overlap the component interval.
'max' uses the exact method to calculate the statistics,
and returns only the maximum identites/positives over
overlapping HSP for the component interval. No averaging
is involved here.
'fast' doesn't involve tiling at all (hence the name),
but it seems like a very good estimate, and uses only
reported values, and so does not require sequence data. It
calculates an average of reported identities, conserved
sites, and lengths, over unmodified hsps in the hit,
weighted by the length of the hsps.
Title : _make_tiling_iterator
Usage : $self->_make_tiling_iterator($type)
Function: Create an iterator code ref that will step through all
minimal combinations of HSPs that produce complete coverage
of the $type ('hit', 'subject', 'query') sequence,
and set the correct iterator property of the invocant
Example :
Returns : The iterator
Args : scalar $type, one of 'hit', 'subject', 'query';
default is 'query'
Title : _tiling_iterator
Usage : $tiling->_tiling_iterator($type,$context)
Function: Retrieve the tiling iterator coderef for the requested
$type ('hit', 'subject', 'query')
Example :
Returns : coderef to the desired iterator
Args : scalar $type, one of 'hit', 'subject', 'query'
default is 'query'
option scalar $context: strand/frame context string
Note : getter only
See also Bio::Search::Tiling::MapTileUtils.
Title : _make_context_key
Alias : _context
Usage : $tiling->_make_context_key(-strand => $strand, -frame => $frame)
Function: create a string indicating strand/frame context; serves as
component of memoizing hash keys
Returns : scalar string
Args : -type => one of ('query', 'hit', 'subject')
-strand => one of (1,0,-1)
-frame => one of (-2, 1, 0, 1, -2)
called w/o args: returns 'all'
Title : _context
Alias : _make_context_key
Usage : $tiling->_make_context_key(-strand => $strand, -frame => $frame)
Function: create a string indicating strand/frame context; serves as
component of memoizing hash keys
Returns : scalar string
Args : -type => one of ('query', 'hit', 'subject')
-strand => one of (1,0,-1)
-frame => one of (-2, 1, 0, 1, -2)
called w/o args: returns 'all'
Most based on a reading of the algorithm name with a configuration lookup.
Title : _contig_intersection
Usage : $tiling->_contig_intersection($type)
Function: Return the minimal set of $type coordinate intervals
covered by the invocant's HSPs
Returns : array of intervals (2-member arrayrefs; see MapTileUtils)
Args : scalar $type: one of 'query', 'hit', 'subject'
Title : _reported_length
Usage : $tiling->_reported_length($type)
Function: Get the total length of the seq $type
for the invocant's hit object, as reported
by (not calculated from) the input data file
Returns : scalar int
Args : scalar $type: one of 'query', 'hit', 'subject'
Note : This is kludgy; the hit object does not currently
maintain accessors for these values, but the
hsps possess these attributes. This is a wrapper
that allows a consistent access method in the
MapTiling code.
Note : Since this number is based on a reported length,
it is already a "logical length".
| BioPerl documentation | Contained in the BioPerl distribution. |
# # BioPerl module for Bio::Search::Tiling::MapTiling # # Please direct questions and support issues to <bioperl-l@bioperl.org> # # Cared for by Mark A. Jensen <maj@fortinbras.us> # # Copyright Mark A. Jensen # # You may distribute this module under the same terms as perl itself # POD documentation - main docs before the code
# Let the code begin... package Bio::Search::Tiling::MapTiling; use strict; use warnings; # Object preamble - inherits from Bio::Root::Root #use lib '../../..'; use Bio::Root::Root; use Bio::Search::Tiling::TilingI; use Bio::Search::Tiling::MapTileUtils; use Bio::LocatableSeq; # use base qw(Bio::Root::Root Bio::Search::Tiling::TilingI); use base qw(Bio::Root::Root Bio::Search::Tiling::TilingI);
sub new { my $class = shift; my @args = @_; my $self = $class->SUPER::new(@args); my($hit, $filter) = $self->_rearrange( [qw( HIT HSP_FILTER)],@args ); $self->throw("HitI object required") unless $hit; $self->throw("Argument must be HitI object") unless ( ref $hit && $hit->isa('Bio::Search::Hit::HitI') ); $self->{hit} = $hit; $self->_set_attributes(); $self->{"_algorithm"} = $hit->algorithm; my @hsps = $hit->hsps; # apply filter function if requested if ( defined $filter ) { if ( ref($filter) eq 'CODE' ) { @hsps = map { $filter->($_) ? $_ : () } @hsps; } else { $self->warn("-filter is not a coderef; ignoring"); } } # identify available contexts for my $t (qw( query hit )) { my %contexts; for my $i (0..$#hsps) { my $ctxt = $self->_context( -type => $t, -strand => $hsps[$i]->strand($t), -frame => $hsps[$i]->frame($t)); $contexts{$ctxt} ||= []; push @{$contexts{$ctxt}}, $i; } $self->{"_contexts_${t}"} = \%contexts; } $self->warn("No HSPs present in hit after filtering") unless (@hsps); $self->hsps(\@hsps); return $self; } # a tiling is based on the set of hsps contained in a single hit. # check all the boundaries - zero hsps, one hsp, all disjoint hsps
sub next_tiling{ my $self = shift; my ($type, $context) = @_; $self->_check_type_arg(\$type); $self->_check_context_arg($type, \$context); return $self->_tiling_iterator($type, $context)->(); }
sub rewind_tilings{ my $self = shift; my ($type,$context) = @_; $self->_check_type_arg(\$type); $self->_check_context_arg($type, \$context); return $self->_tiling_iterator($type, $context)->('REWIND'); }
sub get_tiled_alns { my $self = shift; my ($type, $context) = @_; $self->_check_type_arg(\$type); $self->_check_context_arg($type, \$context); my $t = shift; # first arg after type/context, arrayref to a tiling my @tiling; if ($t && (ref($t) eq 'ARRAY')) { @tiling = @$t; } else { # otherwise, take the first tiling available @tiling = $self->_make_tiling_iterator($type,$context)->(); } my @ret; my @map = $self->coverage_map($type, $context); my @intervals = map {$_->[0]} @map; # disjoint decomp # divide into disjoint covering groups my @groups = covering_groups(@intervals); require Bio::SimpleAlign; require Bio::SeqFeature::Generic; # cut hsp aligns along the disjoint decomp # look for gaps...or add gaps? my ($q_start, $h_start, $q_strand, $h_strand); # build seqs for my $grp (@groups) { my $taln = Bio::SimpleAlign->new(); my (@qfeats, @hfeats); my $query_string = ''; my $hit_string = ''; my ($qlen,$hlen) = (0,0); my ($qinc, $hinc, $qstart, $hstart); for my $intvl (@$grp) { # following just chooses the first available hsp containing the # interval -- this is arbitrary, could be smarter. my $aln = ( containing_hsps($intvl, @tiling) )[0]->get_aln; my $qseq = $aln->get_seq_by_pos(1); my $hseq = $aln->get_seq_by_pos(2); $qstart ||= $qseq->start; $hstart ||= $hseq->start; # calculate the slice boundaries my ($beg, $end); for ($type) { /query/ && do { $beg = $aln->column_from_residue_number($qseq->id, $intvl->[0]); $end = $aln->column_from_residue_number($qseq->id, $intvl->[1]); last; }; /subject|hit/ && do { $beg = $aln->column_from_residue_number($hseq->id, $intvl->[0]); $end = $aln->column_from_residue_number($hseq->id, $intvl->[1]); last; }; } $aln = $aln->slice($beg, $end); $qseq = $aln->get_seq_by_pos(1); $hseq = $aln->get_seq_by_pos(2); $qinc = $qseq->length - $qseq->num_gaps($Bio::LocatableSeq::GAP_SYMBOLS); $hinc = $hseq->length - $hseq->num_gaps($Bio::LocatableSeq::GAP_SYMBOLS); push @qfeats, Bio::SeqFeature::Generic->new( -start => $qlen+1, -end => $qlen+$qinc, -strand => $qseq->strand, -primary => 'query', -source_tag => 'BLAST', -display_name => 'query coordinates', -tag => { query_id => $qseq->id, query_desc => $qseq->desc, query_start => $qstart + (($qseq->strand && $qseq->strand < 0) ? -1 : 1)*$qlen, query_end => $qstart + (($qseq->strand && $qseq->strand < 0) ? -1 : 1)*($qlen+$qinc-1), } ); push @hfeats, Bio::SeqFeature::Generic->new( -start => $hlen+1, -end => $hlen+$hinc, -strand => $hseq->strand, -primary => 'subject/hit', -source_tag => 'BLAST', -display_name => 'subject/hit coordinates', -tag => { subject_id => $hseq->id, subject_desc => $hseq->desc, subject_start => $hstart + (($hseq->strand && $hseq->strand < 0) ? -1 : 1)*$hlen, subject_end => $hstart + (($hseq->strand && $hseq->strand < 0) ? -1 : 1)*($hlen+$hinc-1) } ); $query_string .= $qseq->seq; $hit_string .= $hseq->seq; $qlen += $qinc; $hlen += $hinc; } # create the LocatableSeqs and add the features to each # then add the seqs to the new aln # note in MapTileUtils, Bio::FeatureHolderI methods have been # mixed into Bio::LocatableSeq my $qseq = Bio::LocatableSeq->new( -id => 'query', -seq => $query_string); $qseq->add_SeqFeature(@qfeats); my $hseq = Bio::LocatableSeq->new( -id => 'subject', -seq => $hit_string ); $hseq->add_SeqFeature(@hfeats); $taln->add_seq($qseq); $taln->add_seq($hseq); push @ret, $taln; } return @ret; }
sub identities{ my $self = shift; my ($type, $action, $context) = @_; $self->_check_type_arg(\$type); $self->_check_action_arg(\$action); $self->_check_context_arg($type, \$context); if (!defined $self->{"identities_${type}_${action}_${context}"}) { $self->_calc_stats($type, $action, $context); } return $self->{"identities_${type}_${action}_${context}"}; }
sub conserved{ my $self = shift; my ($type, $action, $context) = @_; $self->_check_type_arg(\$type); $self->_check_action_arg(\$action); $self->_check_context_arg($type, \$context); if (!defined $self->{"conserved_${type}_${action}_${context}"}) { $self->_calc_stats($type, $action, $context); } return $self->{"conserved_${type}_${action}_${context}"}; }
sub length{ my $self = shift; my ($type,$action,$context) = @_; $self->_check_type_arg(\$type); $self->_check_action_arg(\$action); $self->_check_context_arg($type, \$context); if (!defined $self->{"length_${type}_${action}_${context}"}) { $self->_calc_stats($type, $action, $context); } return $self->{"length_${type}_${action}_${context}"}; }
sub frac { my $self = shift; my @args = @_; my ($type, $denom, $action, $context, $method) = $self->_rearrange([qw(TYPE DENOM ACTION CONTEXT METHOD)],@args); $self->_check_type_arg(\$type); $self->_check_action_arg(\$action); $self->_check_context_arg($type, \$context); unless ($method and grep(/^$method$/, qw( identical conserved ))) { $self->throw("-method must specified; one of ('identical', 'conserved')"); } $denom ||= 'total'; unless (grep /^$denom/, qw( total aligned )) { $self->throw("Denominator selection must be one of ('total', 'aligned'), not '$denom'"); } my $key = "frac_${method}_${type}_${denom}_${action}_${context}"; my $stat; for ($method) { $_ eq 'identical' && do { $stat = $self->identities($type, $action, $context); last; }; $_ eq 'conserved' && do { $stat = $self->conserved($type, $action, $context); last; }; do { $self->throw("What are YOU doing here?"); }; } if (!defined $self->{$key}) { for ($denom) { /total/ && do { $self->{$key} = $stat/$self->_reported_length($type); # need fudge fac?? last; }; /aligned/ && do { $self->{$key} = $stat/$self->length($type,$action,$context); last; }; do { $self->throw("What are YOU doing here?"); }; } } return $self->{$key}; }
sub frac_identical{ my $self = shift; my @args = @_; my ($type, $denom, $action,$context) = $self->_rearrange( [qw[ TYPE DENOM ACTION CONTEXT]],@args ); $self->frac( -type=>$type, -denom=>$denom, -action=>$action, -method=>'identical', -context=>$context); }
sub frac_conserved{ my $self = shift; my @args = @_; my ($type, $denom, $action, $context) = $self->_rearrange( [qw[ TYPE DENOM ACTION CONTEXT]],@args ); $self->frac( -type=>$type, -denom=>$denom, -action=>$action, -context=>$context, -method=>'conserved'); }
sub frac_aligned{ my ($self, @args) = @_; my ($type, $action, $context) = $self->_rearrange([qw(TYPE ACTION CONTEXT)],@args); $self->_check_type_arg(\$type); $self->_check_action_arg(\$action); $self->_check_context_arg($type, \$context); if (!$self->{"frac_aligned_${type}_${action}_${context}"}) { $self->{"frac_aligned_${type}_${action}_${context}"} = $self->num_aligned($type,$action,$context)/$self->_reported_length($type); } return $self->{"frac_aligned_${type}_${action}_${context}"}; } sub frac_aligned_query { shift->frac_aligned(-type=>'query', @_) } sub frac_aligned_hit { shift->frac_aligned(-type=>'hit', @_) }
sub num_aligned { shift->length( @_ ) };
sub num_unaligned { my $self = shift; my ($type,$action,$context) = @_; my $ret; $self->_check_type_arg(\$type); $self->_check_action_arg(\$action); $self->_check_context_arg($type, \$context); if (!defined $self->{"num_unaligned_${type}_${action}_${context}"}) { $self->{"num_unaligned_${type}_${action}_${context}"} = $self->_reported_length($type)-$self->num_aligned($type,$action,$context); } return $self->{"num_unaligned_${type}_${action}_${context}"}; }
sub range { my ($self, $type, $context) = @_; $self->_check_type_arg(\$type); $self->_check_context_arg($type, \$context); my @a = $self->_contig_intersection($type,$context); return ($a[0][0], $a[-1][1]); }
sub coverage_map{ my $self = shift; my ($type, $context) = @_; $self->_check_type_arg(\$type); $self->_check_context_arg($type, \$context); if (!defined $self->{"coverage_map_${type}_${context}"}) { # following calculates coverage maps in all strands/frames # if necessary $self->_calc_coverage_map($type, $context); } # if undef is returned, then there were no hsps for given strand/frame if (!defined $self->{"coverage_map_${type}_${context}"}) { $self->warn("No HSPS present for type '$type' in context '$context' for this hit"); return undef; } return @{$self->{"coverage_map_${type}_${context}"}}; }
sub coverage_map_as_text{ my $self = shift; my ($type, $context, $legend_q) = @_; $self->_check_type_arg(\$type); $self->_check_context_arg($type, \$context); my @map = $self->coverage_map($type, $context); my @ret; my @hsps = $self->hit->hsps; my %hsps_i; require Tie::RefHash; tie %hsps_i, 'Tie::RefHash'; @hsps_i{@hsps} = (0..$#hsps); my @mx; foreach (0..$#map) { my @hspx = ('') x @hsps; my @these_hsps = @{$map[$_]->[1]}; @hspx[@hsps_i{@these_hsps}] = ('*') x @these_hsps; $mx[$_] = \@hspx; } untie %hsps_i; push @ret, "\tIntvl\n"; push @ret, "HSPS\t", join ("\t", (0..$#map)), "\n"; foreach my $h (0..$#hsps) { push @ret, join("\t", $h, map { $mx[$_][$h] } (0..$#map) ),"\n"; } if ($legend_q) { push @ret, "Interval legend\n"; foreach (0..$#map) { push @ret, sprintf("%d\t[%d, %d]\n", $_, @{$map[$_][0]}); } push @ret, "HSP legend\n"; my @ints = get_intervals_from_hsps($type,@hsps); foreach (0..$#hsps) { push @ret, sprintf("%d\t[%d, %d]\n", $_, @{$ints[$_]}); } } return @ret; }
sub hit{ my $self = shift; $self->warn("Getter only") if @_; return $self->{'hit'}; }
sub hsps{ my $self = shift; return $self->{'hsps'} = shift if @_; return @{$self->{'hsps'}}; }
sub contexts{ my $self = shift; my ($type, $context) = @_; $self->_check_type_arg(\$type); return keys %{$self->{"_contexts_$type"}} unless defined $context; return undef unless $self->{"_contexts_$type"}{$context}; return @{$self->{"_contexts_$type"}{$context}}; }
sub mapping{ my $self = shift; my $type = shift; $self->_check_type_arg(\$type); return $self->{"_mapping_${type}"}; }
sub default_context{ my $self = shift; my $type = shift; $self->_check_type_arg(\$type); return $self->{"_def_context_${type}"}; }
sub algorithm{ my $self = shift; $self->warn("Getter only") if @_; return $self->{"_algorithm"}; }
sub _calc_coverage_map { my $self = shift; my ($type) = @_; $self->_check_type_arg(\$type); # obtain the [start, end] intervals for all hsps in the hit (relative # to the type) unless ($self->{'hsps'}) { $self->warn("No HSPs for this hit"); return; } my (@map, @hsps, %filters, @intervals); # conversion here? my $c = $self->mapping($type); # create the possible maps for my $context ($self->contexts($type)) { @map = (); @hsps = ($self->hsps)[$self->contexts($type, $context)]; @intervals = get_intervals_from_hsps( $type, @hsps ); # the "frame" my $f = ($intervals[0]->[0] - 1) % $c; # convert interval endpoints... for (@intervals) { $$_[0] = ($$_[0] - $f + $c - 1)/$c; $$_[1] = ($$_[1] - $f)/$c; } # determine the minimal set of disjoint intervals that cover the # set of hsp intervals my @dj_set = interval_tiling(\@intervals); # decompose each disjoint interval into another set of disjoint # intervals, each of which is completely contained within the # original hsp intervals with which it overlaps my $i=0; my @decomp; for my $dj_elt (@dj_set) { my ($covering, $indices) = @$dj_elt; my @covering_hsps = @hsps[@$indices]; my @coverers = @intervals[@$indices]; @decomp = decompose_interval( \@coverers ); for (@decomp) { my ($component, $container_indices) = @{$_}; push @map, [ $component, [@covering_hsps[@$container_indices]] ]; } 1; } # unconvert the components: ##### foreach (@map) { $$_[0][0] = $c*$$_[0][0] - $c + 1 + $f; $$_[0][1] = $c*$$_[0][1] + $f; } foreach (@dj_set) { $$_[0][0] = $c*$$_[0][0] - $c + 1 + $f; $$_[0][1] = $c*$$_[0][1] + $f; } # sort the map on the interval left-ends @map = sort { $a->[0][0]<=>$b->[0][0] } @map; $self->{"coverage_map_${type}_${context}"} = [@map]; # set the _contig_intersection attribute here (side effect) $self->{"_contig_intersection_${type}_${context}"} = [map { $$_[0] } @map]; } return 1; # success }
sub _calc_stats { my $self = shift; my ($type, $action, $context) = @_; # need to check args here, in case method is called internally. $self->_check_type_arg(\$type); $self->_check_action_arg(\$action); $self->_check_context_arg($type, \$context); my ($ident, $cons, $length) = (0,0,0); # fast : avoid coverage map altogether, get a pretty damn # good estimate with a weighted average of reported hsp # statistics ($action eq 'fast') && do { my @hsps = $self->hit->hsps; @hsps = @hsps[$self->contexts($type, $context)]; # weights for averages my @wt = map {$_->length($type)} @hsps; my $sum = eval( join('+',@wt) ); $_ /= $sum for (@wt); for (@hsps) { my $wt = shift @wt; $ident += $wt*$_->matches_MT($type,'identities'); $cons += $wt*$_->matches_MT($type,'conserved'); $length += $wt*$_->length($type); } }; # or, do tiling # calculate identities/conserved sites in tiling # estimate based on the fraction of the component interval covered # and ident/cons reported by the HSPs ($action ne 'fast') && do { foreach ($self->coverage_map($type, $context)) { my ($intvl, $hsps) = @{$_}; my $len = ($$intvl[1]-$$intvl[0]+1); my $ncover = ($action eq 'max') ? 1 : scalar @$hsps; my ($acc_i, $acc_c) = (0,0); foreach my $hsp (@$hsps) { for ($action) { ($_ eq 'est') && do { my ($inc_i, $inc_c) = $hsp->matches_MT( -type => $type, -action => 'searchutils', ); my $frac = $len/$hsp->length($type); $acc_i += $inc_i * $frac; $acc_c += $inc_c * $frac; last; }; ($_ eq 'max') && do { my ($inc_i, $inc_c) = $hsp->matches_MT( -type => $type, -action => 'searchutils', -start => $$intvl[0], -end => $$intvl[1] ); $acc_i = ($acc_i > $inc_i) ? $acc_i : $inc_i; $acc_c = ($acc_c > $inc_c) ? $acc_c : $inc_c; last; }; (!$_ || ($_ eq 'exact')) && do { my ($inc_i, $inc_c) = $hsp->matches_MT( -type => $type, -action => 'searchutils', -start => $$intvl[0], -end => $$intvl[1] ); $acc_i += $inc_i; $acc_c += $inc_c; last; }; } } $ident += ($acc_i/$ncover); $cons += ($acc_c/$ncover); $length += $len; } }; $self->{"identities_${type}_${action}_${context}"} = $ident; $self->{"conserved_${type}_${action}_${context}"} = $cons; $self->{"length_${type}_${action}_${context}"} = $length; return 1; }
# coverage_map is of the form # ( [ $interval, \@containing_hsps ], ... ) # so, for each interval, pick one of the containing hsps, # and return the union of all the picks. # use the combinatorial generating iterator, with # the urns containing the @containing_hsps for each # interval
sub _make_tiling_iterator { ### create the urns my $self = shift; my ($type, $context) = @_; $self->_check_type_arg(\$type); $self->_check_context_arg($type, \$context); # initialize the urns my @urns = map { [0, $$_[1]] } $self->coverage_map($type, $context); my $FINISHED = 0; my $iter = sub { # rewind? if (my $rewind = shift) { # reinitialize urn indices $$_[0] = 0 for (@urns); $FINISHED = 0; return 1; } # check if done... return if $FINISHED; my $finished_incrementing = 0; # @ret is the collector of urn choices my @ret; for my $urn (@urns) { my ($n, $hsps) = @$urn; push @ret, $$hsps[$n]; unless ($finished_incrementing) { if ($n == $#$hsps) { $$urn[0] = 0; } else { ($$urn[0])++; $finished_incrementing = 1 } } } # backstop... $FINISHED = 1 unless $finished_incrementing; # uniquify @ret # $hsp->rank is a unique identifier for an hsp in a hit. # preserve order in @ret my (%order, %uniq); @order{(0..$#ret)} = @ret; $uniq{$order{$_}->rank} = $_ for (0..$#ret); @ret = @order{ sort {$a<=>$b} values %uniq }; return @ret; }; return $iter; }
sub _tiling_iterator { my $self = shift; my ($type, $context) = @_; $self->_check_type_arg(\$type); $self->_check_context_arg($type, \$context); if (!defined $self->{"_tiling_iterator_${type}_${context}"}) { $self->{"_tiling_iterator_${type}_${context}"} = $self->_make_tiling_iterator($type,$context); } return $self->{"_tiling_iterator_${type}_${context}"}; }
sub _check_type_arg { my $self = shift; my $typeref = shift; $$typeref ||= 'query'; $self->throw("Unknown type '$$typeref'") unless grep(/^$$typeref$/, qw( hit query subject )); $$typeref = 'hit' if $$typeref eq 'subject'; return 1; } sub _check_action_arg { my $self = shift; my $actionref = shift; if (!$$actionref) { $$actionref = ($self->_has_sequence_data ? 'exact' : 'est'); } else { $self->throw("Calc action '$$actionref' unrecognized") unless grep /^$$actionref$/, qw( est exact fast max ); if ($$actionref ne 'est' and !$self->_has_sequence_data) { $self->warn("Blast file did not possess sequence data; defaulting to 'est' action"); $$actionref = 'est'; } } return 1; } sub _check_context_arg { my $self = shift; my ($type, $contextref) = @_; if (!$$contextref) { $self->throw("Type '$type' requires strand/frame context for algorithm ".$self->algorithm) unless ($self->mapping($type) == 1); # set default according to default_context attrib $$contextref = $self->default_context($type); } else { ($$contextref =~ /^[mp]$/) && do { $$contextref .= '_' }; $self->throw("Context '$$contextref' unrecognized") unless $$contextref =~ /all|[mp][0-2_]/; } }
sub _make_context_key { my $self = shift; my @args = @_; my ($type, $strand, $frame) = $self->_rearrange([qw(TYPE STRAND FRAME)], @args); _check_type_arg(\$type); return 'all' unless (defined $strand or defined $frame); if ( defined $strand && $self->_has_strand($type) ) { if (defined $frame && $self->_has_frame($type)) { return ($strand >= 0 ? 'p' : 'm').abs($frame); } else { return ($strand >= 0 ? 'p_' : 'm_'); } } else { if (defined $frame && $self->_has_frame($type)) { $self->warn("Frame defined without strand; punting with plus strand"); return 'p'.abs($frame); } else { return 'all'; } } }
sub _context { shift->_make_context_key(@_) }
sub _has_sequence_data { my $self = shift; $self->throw("Hit attribute not yet set") unless defined $self->hit; return (($self->hit->hsps)[0]->seq_str('match') ? 1 : 0); }
sub _has_logical_length { my $self = shift; my $type = shift; $self->_check_type_arg(\$type); # based on mapping coeff $self->throw("Mapping coefficients not yet set") unless defined $self->mapping($type); return ($self->mapping($type) > 1); }
sub _has_strand { my $self = shift; my $type = shift; $self->_check_type_arg(\$type); return $self->{"_has_strand_${type}"}; }
sub _has_frame { my $self = shift; my $type = shift; $self->_check_type_arg(\$type); return $self->{"_has_frame_${type}"}; }
sub _contig_intersection { my $self = shift; my ($type, $context) = @_; $self->_check_type_arg(\$type); $self->_check_context_arg($type, \$context); if (!defined $self->{"_contig_intersection_${type}_${context}"}) { $self->_calc_coverage_map($type); } return @{$self->{"_contig_intersection_${type}_${context}"}}; }
sub _reported_length { my $self = shift; my $type = shift; $self->_check_type_arg(\$type); my $key = uc( $type."_LENGTH" ); return ($self->hsps)[0]->{$key}; } 1;