def _blast_feature(self, f, c1, c2):
trans = Translator(self._abort_event)
cds = trans.translate(f.extract(c1), 11)
sixframes = trans.translate_six_frames_single(c2, 11)
if not sixframes: return [(None, None, None)]
results = []
for frame in sixframes:
res = BlastCLI.s2s_blast(cds, frame, self.evalue, command='blastp', task='blastp')
if res: results.extend(res)
hsps = BlastCLI.all_hsps(results)
if not hsps: return [(None, None, None)]
f1 = []
f2 = []
col = []
fname = self._feature_name(f, default='CDS')
cds_len = len(cds)
min_len = len(cds) * self.min_length
for hsp in hsps:
if hsp.align_length < min_len: continue
if hsp.identities / float(hsp.align_length) < self.min_identity: continue
color_t = (float(hsp.identities) / hsp.align_length)
print '%s %s: %5.1f%% (%5.1f%%)' % (c1.description, fname, color_t * 100, float(hsp.identities) / cds_len * 100)
col.append(colors.linearlyInterpolatedColor(colors.Color(0, 0, 1, 0.2), colors.Color(0, 1, 0, 0.2),
0.2, 1, color_t))
qstart = (hsp.query_start - 1) * 3
qend = qstart + hsp.align_length * 3
sstart = (hsp.sbjct_start - 1) * 3
send = sstart + hsp.align_length * 3
f1.append(
SeqFeature(FeatureLocation(f.location.start + qstart, f.location.start + qend, strand=hsp.strand[0])))
f2.append(SeqFeature(FeatureLocation(sstart, send, strand=hsp.strand[1])))
return zip(f1, f2, col)
def hmmsearch_genes(self,
hmms,
genome,
table='Standard',
decorate=False,
**kwargs):
#get _genes
genes = get_indexes_of_all_genes(genome)
if not genes: return None
for gene_id, gi in enumerate(genes):
genome.features[gi].qualifiers['feature_id'] = gi
genome.features[gi].qualifiers['gene_id'] = gene_id
#translate _genes
with user_message('Translating _genes/CDS of %s' % genome.description,
'\n'):
translator = Translator(self._abort_event)
translation = translator.translate_features(genome, genes, table)
if not translation: return None
if isinstance(hmms, str): hmms = [hmms]
results = dict()
for hmm in hmms:
with user_message('Performing hmm search.'):
hmm_results = self.hmmsearch_recs(hmm, translation, **kwargs)
if not hmm_results: return None
with user_message('Parsing search results...'):
#get hit_ids of hmm matches
hits = dict()
for result in hmm_results:
for hit in result.iterhits():
hits[hit.id] = hit
#get indexes of features where hmm hit
hit_features = dict()
for t in translation:
if t.id in hits:
fid = t.features[0].qualifiers.get('feature_id')
if fid is None: continue
hit_features[fid] = hits[t.id], t
if hit_features: results.update(hit_features)
#decorate genome
if decorate:
with user_message('Adding results as annotations...'):
hmm_name = os.path.basename(hmm)
for f in hit_features:
feature = genome.features[f]
for hsp in hit_features[f][0]:
if feature.strand == 1:
hmm_location = FeatureLocation(
feature.location.start + hsp.hit_start * 3,
feature.location.start + hsp.hit_end * 3,
feature.strand)
else:
hmm_location = FeatureLocation(
feature.location.end - hsp.hit_end * 3,
feature.location.end - hsp.hit_start * 3,
feature.strand)
hmm_feature = self.hsp2feature(
hmm_name, 'HMM_annotations', hmm_location, hsp)
genome.features.append(hmm_feature)
return results if results else None
def hmmsearch_genome(self, hmm, genome, table='Standard', decorate=False, **kwargs):
#get genes
genes = get_indexes_of_genes(genome)
if not genes: return None
for gene_id, gi in enumerate(genes):
genome.features[gi].qualifiers['feature_id'] = gi
genome.features[gi].qualifiers['gene_id'] = gene_id
#translate genes
with user_message('Translating genes/CDS of %s' % genome.description, '\n'):
translator = Translator(self._abort_event)
translation = translator.translate(genome, genes, table)
if not translation: return None
with user_message('Performing hmm search.'):
results = self.hmmsearch_recs(hmm, translation)
if not results: return None
with user_message('Parsing search results...'):
#get hit_ids of hmm matches
hits = dict()
for result in results:
for hit in result.iterhits():
hits[hit.id] = hit
#get indexes of features where hmm hit
hit_features = dict()
for t in translation:
if t.id in hits:
fid = t.features[0].qualifiers.get('feature_id')
if fid is None: continue
hit_features[fid] = hits[t.id], t
#decorate genome
if decorate:
with user_message('Adding results as annotations...'):
hmm_name = os.path.basename(hmm)
for f in hit_features:
feature = genome.features[f]
for hsp in hit_features[f][0]:
if feature.strand == 1:
hmm_location = FeatureLocation(feature.location.start+hsp.hit_start*3,
feature.location.start+hsp.hit_end*3,
feature.strand)
else:
hmm_location = FeatureLocation(feature.location.end-hsp.hit_end*3,
feature.location.end-hsp.hit_start*3,
feature.strand)
hmm_feature = SeqFeature(hmm_location, type='misc_feature')
hmm_feature.qualifiers['hmm_model'] = hmm_name
hmm_feature.qualifiers['bitscore'] = hsp.bitscore
hmm_feature.qualifiers['psi_evalue'] = hsp.psi_evalue
hmm_feature.qualifiers['evalue_cond'] = hsp.evalue_cond
hmm_feature.qualifiers['acc_average'] = hsp.acc_avg
hmm_feature.qualifiers['bias'] = hsp.bias
genome.features.append(hmm_feature)
print 'Done.\n'
return hit_features
def _blast_feature(self, f, c1, c2, features1, features2, evalue,
max_rlen):
trans = Translator(self._abort_event)
cds = trans.translate(f.extract(c1), 11)
sixframes = trans.translate_six_frames_single(c2, 11)
if not sixframes: return [(None, None, None)]
results = []
for frame in sixframes:
res = BlastCLI.s2s_blast(cds,
frame,
evalue,
command='blastp',
task='blastp')
if res: results.extend(res)
hsps = BlastCLI.all_hsps(results, max_rlen)
if not hsps: return [(None, None, None)]
f1 = []
f2 = []
col = []
c1_name = pretty_rec_name(c1)
if 'locus_tag' in f.qualifiers:
fname = f.qualifiers['locus_tag'][0]
else:
fname = 'CDS'
cds_len = len(cds)
for hsp in hsps:
color_t = (float(hsp.identities) / hsp.align_length)
print '%s %s: %5.1f%% (%5.1f%%)' % (c1_name, fname, color_t * 100,
float(hsp.identities) /
cds_len * 100)
col.append(
colors.linearlyInterpolatedColor(colors.Color(0, 0, 1, 0.2),
colors.Color(0, 1, 0, 0.2),
0.2, 1, color_t))
qstart = (hsp.query_start - 1) * 3
qend = qstart + hsp.align_length * 3
sstart = (hsp.sbjct_start - 1) * 3
send = sstart + hsp.align_length * 3
f1.append(
SeqFeature(
FeatureLocation(f.location.start + qstart,
f.location.start + qend,
strand=hsp.strand[0])))
f2.append(
SeqFeature(FeatureLocation(sstart, send,
strand=hsp.strand[1])))
return zip(f1, f2, col)
def hmmsearch_genome(self,
hmms,
genome,
table='Standard',
decorate=False,
**kwargs):
#translate _genes
with user_message('Translating whole genome in 6 reading frames',
'\n'):
translator = Translator(self._abort_event)
translation = translator.translate_six_frames(genome, table)
if not translation: return None
if isinstance(hmms, str): hmms = [hmms]
results = []
for hmm in hmms:
with user_message('Performing hmm search.'):
hmm_results = self.hmmsearch_recs(hmm, translation, **kwargs)
if not any(len(r) for r in hmm_results): continue
results += hmm_results
#decorate genome
if decorate:
translation = dict((t.id, t) for t in translation)
with user_message('Adding results as annotations...'):
hmm_name = os.path.basename(hmm)
glen = len(genome)
for frame in hmm_results:
for hit in frame:
frec = translation[hit.id]
start = frec.annotations['start']
strand = frec.annotations['strand']
for hsp in hit:
if strand == 1:
hmm_location = FeatureLocation(
start + hsp.hit_start * 3,
start + hsp.hit_end * 3, strand)
else:
hmm_location = FeatureLocation(
glen - start - hsp.hit_end * 3,
glen - start - hsp.hit_start * 3,
strand)
hmm_feature = self.hsp2feature(
hmm_name, 'HMM_annotations', hmm_location,
hsp)
genome.features.append(hmm_feature)
return results if results else None
def blastp_annotate(self, tag_sequences, subject_record, min_identity, evalue=0.001, table=11, **kwargs):
# translate subject in six frames
with user_message('Translating whole genome in 6 reading frames', '\n'):
translator = Translator(self._abort_event)
translation = translator.translate_six_frames(subject_record, table)
if not translation: return False
results = self.s2s_blast_batch(tag_sequences, translation, evalue=evalue, command='blastp', **kwargs)
if results is None: return False
with user_message('Adding results as annotations...'):
annotated = False
subj_len = len(subject_record)
for i, tag in enumerate(tag_sequences):
if not results[i]: continue
tag_name = pretty_rec_name(tag)
if tag_name != tag.id:
tag_name += ' (%s)' % tag.id
for frame, record in enumerate(results[i]):
if not record: continue
frec = translation[frame]
start = frec.annotations['start']
strand = frec.annotations['strand']
for hit in record:
for ali in hit.alignments:
for hsp in ali.hsps:
if hsp.identities / float(hsp.align_length) < min_identity: continue
if strand == 1:
location = FeatureLocation(start+(hsp.sbjct_start-1)*3,
start+hsp.sbjct_end*3,
strand)
else:
location = FeatureLocation(subj_len-start-hsp.sbjct_end*3,
subj_len-start-hsp.sbjct_start*3,
strand)
feature = self.hsp2feature(tag_name, 'blastp_annotations', location, hsp)
self.add_program(feature, 'blastp')
subject_record.features.append(feature)
annotated = True
return annotated
def g2g_blastp(self, reference, subjects, table='Standard',
evalue=0.001, max_rlen=0, features_of_interest=None):
'''
Perform blastp of each coding sequence of the reference against each
subject, which is first translated gene-by-gene.
Parameters
@param reference: SeqRecord object of the reference genome
@param subjects: a list of SeqRecord objects of subject genomes
@param table: translation table number (see NCBI site for description)
@param evalue: filter out blastp results with E-value grater than this
@param max_rlen: filter out blastp results which are shorter than this
fraction of target gene length
@param features_of_interest: list of dictionaries of the form
{qualifier_name : qualifier_value}
to mark features denoting known clusters that should be analyzed one
against the other
@return: list of pairs (CDS, (blast_result1, blast_result2, ...))
where CDS is a gene/CDS feature from the reference.features list
and blast_resultN is a list of results for the N-th
subject, containing following information:
(hit_feature, align_length, percent_identity, evalue)
where hit_feature is a SeqFeature object of the gene/CDS of the subject
where top blast hit is located, align_length is the length of the hit,
percent_identity is the ratio of number of identities and align_length [0; 1]
and evalue is the E-value of the top hit.
'''
if not reference or not subjects:
print 'No reference or subject sequences provided'
return None
#get list of features to query
with user_message('Searching for gene/CDS features in provided sequences...'):
all_records = [reference]+subjects
num_records = len(all_records)
features = self.parallelize_work(1, lambda ri, records: self._get_genes(records[ri]),
range(num_records),
all_records)
if self.aborted():
print '\nAborted'
return None
if not features or not features[0]:
print ('\nReference sequence does not contain annotated genes:\n%s %s'
% (reference.id, reference.description))
return None
if len([f for f in features if f]) < 2:
print '\nSubject sequences do not contain annotated genes'
return None
#add gene ids
for ri, genes in enumerate(features):
if not genes: continue
r = all_records[ri]
for gene_id, gi in enumerate(genes):
r.features[gi].qualifiers['feature_id'] = gi
r.features[gi].qualifiers['gene_id'] = gene_id
#get features of interest if requested
fois = None
if features_of_interest:
with user_message('Searching for features of interest...'):
fois = []
for foi in features_of_interest:
foi = self._get_fois(all_records, foi)
if foi and foi[0]: fois.append(foi)
if self.aborted():
print '\nAborted'
return None
#translate features to proteins
with Progress('Translating genes found in the reference and subjects...', num_records) as prg:
translator = Translator(self._abort_event)
translations = [None]*num_records
foi_translations = [[None]*num_records for _f in fois]
for i, (f, rec) in enumerate(zip(features, all_records)):
if not f:
prg.step(i)
continue
translation = translator.translate(rec, f, table)
if not translation: return None
if i > 0:
translations[i] = cat_records(translation)
if fois:
for ifoi, foi in enumerate(fois):
foi_loc = [0, 0]
for foi_var in foi[i]:
if not foi_var: continue
for gid in foi_var:
l = translations[i].features[gid].location
foi_loc[0] = min(int(l.start)+1, foi_loc[0]) if foi_loc[0] > 0 else int(l.start)+1
foi_loc[1] = max(int(l.end), foi_loc[1])
if foi_loc[0] > 0: foi_translations[ifoi][i] = foi_loc
else:
translations[i] = translation
if fois:
for ifoi, foi in enumerate(fois):
foi_translations[ifoi][i] = [[translation[gid] for gid in foi_var] for foi_var in foi[i]]
prg.step(i)
#blast features against subjects
with user_message('Performing local blast of every translated gene in the reference against every translated subject...', '\n'):
stranslations = translations[1:]
blast_results = self._s2s_blast_batch(translations[0], stranslations, None, evalue,
command='blastp', task='blastp')
if self.aborted():
print '\nAborted'
return None
if not blast_results:
print '\nBlast have not returned any results.'
return None
if fois: #redo blast for fois and replace the results
with user_message('Rerunning blast for FOIs...', '\n'):
for ifoi, foi in enumerate(foi_translations):
sfoi_locs = foi[1:]
for i, foi_var in enumerate(foi[0]):
foi_blast = self._s2s_blast_batch(foi_var, stranslations, sfoi_locs, evalue,
command='blastp', task='blastp')
if self.aborted():
print '\nAborted'
return None
if not foi_blast: continue
for gi, gid in enumerate(fois[ifoi][0][i]):
if foi_blast[gi]:
blast_results[gid] = foi_blast[gi]
#process blast results
pairs = list(itertools.product(xrange(len(translations[0])), xrange(len(stranslations))))
with ProgressCounter('Searching for genes in subjects that overlap with top blast hits...', len(pairs)) as prg:
work = self.Work()
work.start_work(self._find_features_by_hsps, pairs,
None, stranslations, blast_results)
@MultiprocessingBase.results_assembler
def assembler(index, result, blast_results, pairs, prg):
qs = pairs[index]
blast_results[qs[0]][qs[1]] = result
prg.count()
work.assemble(assembler, blast_results, pairs, prg)
if not work.wait(): return None
return zip((reference.features[f] for f in features[0]), blast_results)
def g2g_blastp(self, reference, subjects, table='Standard',
evalue=0.001, max_rlen=0, features_of_interest=None):
'''
Perform blastp of each coding sequence of the reference against each
subject, which is first translated gene-by-gene.
Parameters
@param reference: SeqRecord object of the reference genome
@param subjects: a list of SeqRecord objects of subject genomes
@param table: translation table number (see NCBI site for description)
@param evalue: filter out blastp results with E-value grater than this
@param max_rlen: filter out blastp results which are shorter than this
fraction of target gene length
@param features_of_interest: list of dictionaries of the form
{qualifier_name : qualifier_value}
to mark features denoting known clusters that should be analyzed one
against the other
@return: list of pairs (CDS, (blast_result1, blast_result2, ...))
where CDS is a gene/CDS feature from the reference.features list
and blast_resultN is a list of results for the N-th
subject, containing following information:
(hit_feature, align_length, percent_identity, evalue)
where hit_feature is a SeqFeature object of the gene/CDS of the subject
where top blast hit is located, align_length is the length of the hit,
percent_identity is the ratio of number of identities and align_length [0; 1]
and evalue is the E-value of the top hit.
'''
if not reference or not subjects:
print 'No reference or subject sequences provided'
return None
#get list of features to query
with user_message('Searching for gene/CDS features in provided sequences...'):
all_records = [reference]+subjects
num_records = len(all_records)
features = self.parallelize_work(1, lambda ri, records: self._get_genes(records[ri]),
range(num_records),
all_records)
if self.aborted():
print '\nAborted'
return None
if not features or not features[0]:
print ('\nReference sequence does not contain annotated _genes:\n%s %s'
% (reference.id, reference.description))
return None
if len([f for f in features if f]) < 2:
print '\nSubject sequences do not contain annotated _genes'
return None
#add gene ids
for ri, genes in enumerate(features):
if not genes: continue
r = all_records[ri]
for gene_id, gi in enumerate(genes):
r.features[gi].qualifiers['feature_id'] = gi
r.features[gi].qualifiers['gene_id'] = gene_id
#get features of interest if requested
fois = None
if features_of_interest:
with user_message('Searching for features of interest...'):
fois = []
for foi in features_of_interest:
foi = self._get_fois(all_records, foi)
if foi and foi[0]: fois.append(foi)
if self.aborted():
print '\nAborted'
return None
#translate features to proteins
with Progress('Translating _genes found in the reference and subjects...', num_records) as prg:
translator = Translator(self._abort_event)
translations = [None]*num_records
foi_translations = [[None]*num_records for _f in fois]
for i, (f, rec) in enumerate(zip(features, all_records)):
if not f:
prg.step(i)
continue
translation = translator.translate_features(rec, f, table)
if not translation: return None
if i > 0:
translations[i] = cat_records(translation)
if fois:
for ifoi, foi in enumerate(fois):
foi_loc = [0, 0]
for foi_var in foi[i]:
if not foi_var: continue
for gid in foi_var:
l = translations[i].features[gid].location
foi_loc[0] = min(int(l.start)+1, foi_loc[0]) if foi_loc[0] > 0 else int(l.start)+1
foi_loc[1] = max(int(l.end), foi_loc[1])
if foi_loc[0] > 0: foi_translations[ifoi][i] = foi_loc
else:
translations[i] = translation
if fois:
for ifoi, foi in enumerate(fois):
foi_translations[ifoi][i] = [[translation[gid] for gid in foi_var] for foi_var in foi[i]]
prg.step(i)
#blast features against subjects
with user_message('Performing local blast of every translated gene in the reference against every translated subject...', '\n'):
stranslations = translations[1:]
blast_results = self._s2s_blast_batch(translations[0], stranslations, None, evalue,
command='blastp', task='blastp')
if self.aborted():
print '\nAborted'
return None
if not blast_results:
print '\nBlast have not returned any results.'
return None
if fois: #redo blast for fois and replace the results
with user_message('Rerunning blast for FOIs...', '\n'):
for ifoi, foi in enumerate(foi_translations):
sfoi_locs = foi[1:]
for i, foi_var in enumerate(foi[0]):
foi_blast = self._s2s_blast_batch(foi_var, stranslations, sfoi_locs, evalue,
command='blastp', task='blastp')
if self.aborted():
print '\nAborted'
return None
if not foi_blast: continue
for gi, gid in enumerate(fois[ifoi][0][i]):
if foi_blast[gi]:
blast_results[gid] = foi_blast[gi]
#process blast results
pairs = list(itertools.product(xrange(len(translations[0])), xrange(len(stranslations))))
with ProgressCounter('Searching for _genes in subjects that overlap with top blast hits...', len(pairs)) as prg:
work = self.Work()
work.start_work(self._find_features_by_hsps, pairs,
None, stranslations, blast_results)
@MultiprocessingBase.results_assembler
def assembler(index, result, blast_results, pairs, prg):
qs = pairs[index]
blast_results[qs[0]][qs[1]] = result
prg.count()
work.assemble(assembler, blast_results, pairs, prg)
if not work.wait(): return None
return zip((reference.features[f] for f in features[0]), blast_results)
