def get_reverse_cbg(cbg,frame,verbose=False): """ Get the ReversecomplementCodingBlockGraph in requested frame of this CBG @type cbg: CodingBlockGraph @param cbg: CodingBlockGraph to reversecomplement @type frame: integer @param frame: 0,1 or 2 @type verbose: Boolean @param verbose: print intermediate info to STDOUT for debugging purposes @rtype: ReversecomplementCodingBlockGraph or None @return: ReversecomplementCodingBlockGraph (when existing) or None """ min_orf_length = (cbg.omsrlength()/2)*3 orfs = get_reverse_strand_orfsets(cbg,frame,min_orf_length=min_orf_length) # remap the identifiers of the orf objects i.o.t.... multifastas = {} blastdbs = {} pacbpcol = PacbpCollectionGraph() dpcpacbpcol = PacbpCollectionGraph() # ``deepcopied`` variant for pacbps for org in orfs.keys(): fname = "%s_reversecbg_%s.mfa" % (org,cbg.barcode()) writeMultiFasta(orfs[org].tofastadict(),fname) multifastas[org] = fname ######################################################################## if verbose: print "ORFS:", org, len(orfs[org].orfs), print [len(o.protein_sequence) for o in orfs[org].orfs ] ######################################################################## revpacbps = {} for orgQ,orgS in cbg.pairwisecrosscombinations_organism(): # create blastdb if it does not exist yet if not blastdbs.has_key(orgS): formatdb(fname=multifastas[orgS]) blastdbs[orgS] = multifastas[orgS] revpacbporfs = {} for orfQ in orfs[orgQ].orfs: # run blast_seqs2db blastrec = blastall_seq2db(orfQ.id,orfQ.protein_sequence, dbname="./"+blastdbs[orgS]) if len(blastrec.alignments) == 0: continue for alignment in blastrec.alignments: # obtain coordinates from sbjct orf identifier orfS = orfs[orgS].get_orf_by_id(alignment.title.replace(">","")) # take only the *best* HSP (highest scoring first one) hsp = alignment.hsps[0] # skip if hsp is very short if len(hsp.query) < cbg.omsrlength()/2: continue # correct to absolute positions hsp.query_start = hsp.query_start + orfQ.protein_startPY hsp.sbjct_start = hsp.sbjct_start + orfS.protein_startPY # initialize the PacbP pacbporf = pacb.conversion.pacbp2pacbporf( pacb.PacbP(blastp_hsp=hsp),orfQ,orfS) ################################################################ if verbose: print pacbporf, orgQ,orgS, orfQ print pacbporf.query print pacbporf.match print pacbporf.sbjct ###pacbporf.print_protein_and_dna() ################################################################ nodeQ = ( orgQ, orfQ.protein_startPY ) nodeS = ( orgS, orfS.protein_startPY ) uqkey = pacbporf.construct_unique_key(nodeQ,nodeS) if not nodeQ in pacbpcol.get_nodes(): pacbpcol.add_node(nodeQ) if not nodeS in pacbpcol.get_nodes(): pacbpcol.add_node(nodeS) pacbpcol.add_edge(nodeQ,nodeS,wt=pacbporf.bitscore) # store to dpcpacbpcol -> pacbpcol is broken in pieces lateron! dpcpacbpcol.pacbps[(uqkey,nodeQ,nodeS)] = pacbporf # file cleanup _file_cleanup(multifastas.values()) _file_cleanup(["formatdb.log"]) _file_cleanup([ fname+".*" for fname in blastdbs.values()]) if not pacbpcol.organism_set_size() == cbg.organism_set_size(): # no CBG on the reverse strand return None # ``deepcopy`` PacbPcollection dpcpacbpcol.add_nodes( pacbpcol.get_nodes() ) for (uqkey,nodeQ,nodeS) in dpcpacbpcol.pacbps.keys(): (bitscore,length,orfQid,orfSid) = uqkey dpcpacbpcol.add_edge(nodeQ,nodeS,wt=bitscore) ############################################################################ if verbose: print pacbpcol, "bitscores:", print [ pacbporf.bitscore for pacbporf in dpcpacbpcol.pacbps.values() ] ############################################################################ # do some transformations on the pacbpcol pacbpcol.remove_low_connectivity_nodes(min_connectivity=cbg.node_count()-1) splittedCBGs = pacbpcol.find_fully_connected_subgraphs( edges=cbg.node_count()-1 , max_missing_edges=0 ) # convert to list of CBGs and do some transformations cbgList = ListOfCodingBlockGraphs(splittedCBGs,input={},crossdata={}) cbgList.remove_all_but_complete_cbgs() cbgList.harvest_pacbps_from_pacbpcollection(dpcpacbpcol) cbgList.remove_cbgs_without_omsr() cbgList.update_edge_weights_by_minimal_spanning_range() cbgList.order_list_by_attribute(order_by='total_weight',reversed=True) ############################################################################ if verbose: for revcbg in cbgList: print "revCBG:", revcbg ############################################################################ if not cbgList: # no CBG on the reverse strand return None else: # return the highest scoring CBG as a ReversecomlementCodingBlockGraph return CodingBlockGraph2ReversecomlementCodingBlockGraph( cbgList.codingblockgraphs[0])
def _blastorfset2blastdb(geneQ, geneS, blastdbfname, input, crossdata, GSgraph, blastoptions=None, elegiable_orfsQ=[], elegiable_orfsS_ids=[], logging=False): """ """ hitcnt = 0 for orfQ in elegiable_orfsQ: # check if protein sequence present in Orf object # in obscure cases of unigenes, no protein sequence present! if not orfQ.protein_sequence: continue # make unique node identifier and blast header nodeQ = (geneQ, orfQ.id) header = "%s_orf_%s" % (geneQ, orfQ.id) # do the blastp! blastrec = blastall_seq2db( header, orfQ.protein_sequence, dbname=blastdbfname, extra_blastp_params=blastoptions.extra_blastp_params) # check if blast failed (then, blastrec == False) if not blastrec: continue # check if there are any hits/hsps! if len(blastrec.alignments) == 0: # no hits; continue continue for alignment in blastrec.alignments: # get back orfpointerB from the SBJCT and create nodeS _parts = alignment.title.split("_") geneS = "_".join(_parts[0:-2]).replace('>', '') _orfpointerS = int(_parts[-1]) nodeS = (geneS, _orfpointerS) # ignore hit if nodeS orfid not occurring in the NON-empty list elegiable_orfsS_ids if elegiable_orfsS_ids and _orfpointerS not in elegiable_orfsS_ids: continue # get the Orf object of this sbjct sequence orfS = input[geneS]['orfs'].get_orf_by_id(_orfpointerS) # loop over the HSPs for hsp in alignment.hsps: # If hits are really tiny (happens in case of BLOSUM45 matrix), # discard them directly before precious time is lost... if len(hsp.query ) <= blastoptions.BLASTP_DIRECTLY_IGNORE_TINY_HITS: continue # correct to absolute positions hsp.query_start = hsp.query_start + orfQ.protein_startPY hsp.sbjct_start = hsp.sbjct_start + orfS.protein_startPY hsp.query_end = hsp.query_end + orfQ.protein_startPY hsp.sbjct_end = hsp.sbjct_end + orfS.protein_startPY # VERY exceptional case: HSP starts or ends with a gap # I expect this is an error in Blastp .... strip_exterior_gaps(hsp) if hsp.query.find(" ") > 0: # VERY exceptional case: erroneously NCBI parsed HSP: # Score 8 (7 bits), expectation 1.7e+01, alignment length 41 # Query: 1622 STHTYDAC TRCI----PFVDTGHKHENPTEALLDSTA 1654 # TR P + H + P+++ S++ # Sbjct: 489 TEHIYLHT TRSTWPPKPPTNASHANTKPSKSHHRSSS 525 if len(hsp.query.split(" ")[-1]) == len(hsp.match): hsp.query = hsp.query.split(" ")[-1] hsp.sbjct = hsp.sbjct.split(" ")[-1] elif len(hsp.query.split(" ")[0]) == len(hsp.match): hsp.query = hsp.query.split(" ")[0] hsp.sbjct = hsp.sbjct.split(" ")[0] elif len(hsp.query) == len(hsp.match): # spaces in both query/match/sbjct while hsp.query.find(" ") > 0: pos = hsp.query.find(" ") if hsp.sbjct[pos] == " " and hsp.match[pos] == " ": hsp.query = hsp.query[0:pos] + hsp.query[pos + 1:] hsp.match = hsp.match[0:pos] + hsp.match[pos + 1:] hsp.sbjct = hsp.sbjct[0:pos] + hsp.sbjct[pos + 1:] else: # HPS is not repairable -> quit trying break elif len(hsp.query) == len(hsp.sbjct): while hsp.query.find(" ") > 0: pos = hsp.query.find(" ") hsp.query = hsp.query[0:pos] + hsp.query[pos + 1:] hsp.sbjct = hsp.sbjct[0:pos] + hsp.sbjct[pos + 1:] # recreate alignment match string is done upon # creation of PacbP object else: pass # VERY exceptional case: HSP starts or ends with a gap # I expect this is an error in Blastp .... strip_exterior_gaps(hsp) try: pacbp = pacb.PacbP(blastp_hsp=hsp, MATRIX=blastoptions.MATRIX) except: # VERY exceptional miscelaneous cases: erroneously NCBI parsed HSP: print hsp print "'%s' X" % hsp.query, len( hsp.query), hsp.query_start, hsp.query_end print "'%s' X" % hsp.match, len(hsp.match) print "'%s' X" % hsp.sbjct, len( hsp.sbjct), hsp.sbjct_start, hsp.sbjct_end pacbp = pacb.PacbP(blastp_hsp=hsp, MATRIX=blastoptions.MATRIX) # make pacbp of this hsp pacbp = pacb.PacbP(blastp_hsp=hsp, MATRIX=blastoptions.MATRIX) # if logging is requested for, print this pacbp to STDOUT if logging: print ">>> Q", nodeQ, orfQ.tcode_symbolic( ), "S", nodeS, orfS.tcode_symbolic( ), pacbp, blastoptions.MATRIX.name, hsp.expect, hsp.bits print ">>>", blastoptions.extra_blastp_params if pacbp.length > 100: print pacbp.query[0:40] + '.' * 7 + str( pacbp.length - 80) + '.' * 7 + pacbp.query[-40:] print pacbp.match[0:40] + '.' * 7 + str( pacbp.length - 80) + '.' * 7 + pacbp.match[-40:] print pacbp.sbjct[0:40] + '.' * 7 + str( pacbp.length - 80) + '.' * 7 + pacbp.sbjct[-40:] else: print pacbp.query print pacbp.match print pacbp.sbjct # blastoptions.BLASTP_HSP_MINIMAL_LENGTH represents the minimal # length of the aligned part. (To) short pacbp's are abandoned if pacbp.length < blastoptions.BLASTP_HSP_MINIMAL_LENGTH: if pacbp.identityscore == float(pacbp.length): # escape for 100% identical tiny pacbps pass elif pacbp.identity + pacbp.similarity == pacbp.length: # escape for 100% similar tiny pacbps pass else: # pacbp is to small. Discard! if logging: print "to small..." continue # check if the pacbp is not conflicting with the currect GSG graph # if so, ignore now because it will not yield a proper edge in an (accepted) CBG! if GSgraph and len( GSgraph ) and GSgraph.is_pacbp_conflicting_with_genestructure( pacbp, orgQ=geneQ, orgS=geneS): ###print "GSGconflict!", nodeQ,nodeS, GSgraph.is_pacbp_conflicting_with_genestructure(pacbp,orgQ=geneQ,orgS=geneS), len(pacbp) continue # here we have a potentially accepted pacbp. # make a/the unique key of this pacbp key = (pacbp.bits, pacbp.length, orfQ.id, _orfpointerS) # check for evalue criterion if (blastoptions.BLASTP_HSP_MAXIMAL_EXPECT or blastoptions.BLASTP_HSP_MAXIMAL_EXPECT == 0.0 ) and hsp.expect > blastoptions.BLASTP_HSP_MAXIMAL_EXPECT: # pacbp is long enough but has a to high evalue crossdata[(geneQ, geneS)]['lowscoring_pacbs'][key] = pacbp if logging: print "to low bitscore or expect" continue # check for bitscore criterion if (blastoptions.BLASTP_HSP_MINIMAL_BITS or blastoptions.BLASTP_HSP_MINIMAL_BITS == 0 ) and pacbp.bits < blastoptions.BLASTP_HSP_MINIMAL_BITS: # pacbp is long enough but has a to low bitscore crossdata[(geneQ, geneS)]['lowscoring_pacbs'][key] = pacbp if logging: print "to low bitscore or expect" continue # !!Hurray!! an accepted pacbp. Store to crossdata # store it to the 'accepted_pacbs' dict of crossdata crossdata[(geneQ, geneS)]['accepted_pacbs'][key] = pacbp hitcnt += 1 if logging: print "ACCEPTED" # done -> check next orf! # return the filled crossdata structure return crossdata, hitcnt
def get_frameshifted_cbg(cbg, input, verbose=True): """ Get a CBG with frameshifts (in some of if Orfs) compared to this CBG @type cbg: CodingBlockGraph @param cbg: CodingBlockGraph to check for frameshifts @type input: dict @param input: input <dict data structure> with lists of Orfs @type verbose: Boolean @param verbose: print intermediate info to STDOUT for debugging purposes @rtype: CodingBlockGraph or None @return: CodingBlockGraph (when existing) or None """ # get elegiable lists of Orfs orfs = _get_elegiable_frameshift_orfsets(cbg, input) # check how many Orfs are elgiable... if sum([len(l.orfs) for l in orfs.values()]) == cbg.node_count(): # no frameshift possible here... return None # remap the identifiers of the orf objects i.o.t.... multifastas = {} blastdbs = {} pacbpcol = PacbpCollectionGraph() dpcpacbpcol = PacbpCollectionGraph() # ``deepcopied`` variant for pacbps for org in orfs.keys(): # REMAP fastaheaders as ids to retrieve the Orfs after blast.. for orf in orfs[org].orfs: orf.fastaheader = str(orf.id) fname = "%s_frameshiftcbg_%s.mfa" % (org, cbg.barcode()) writeMultiFasta(orfs[org].tofastadict(), fname) multifastas[org] = fname ######################################################################## if verbose: print "ORFS:", org, len(orfs[org].orfs), print[orf.id for orf in orfs[org].orfs], print[str(orf) for orf in orfs[org].orfs] ######################################################################## for orgQ, orgS in cbg.pairwisecrosscombinations_organism(): # create blastdb if it does not exist yet if not blastdbs.has_key(orgS): formatdb(fname=multifastas[orgS]) blastdbs[orgS] = multifastas[orgS] for orfQ in orfs[orgQ].orfs: # run blast_seqs2db blastrec = blastall_seq2db(orfQ.id, orfQ.protein_sequence, dbname="./" + blastdbs[orgS]) if len(blastrec.alignments) == 0: continue for alignment in blastrec.alignments: # obtain coordinates from sbjct orf identifier orfid = alignment.title.replace(">", "").split(" ")[0].replace( "_", "") orfS = orfs[orgS].get_orf_by_id(int(orfid)) nodeQ = (orgQ, orfQ.id) nodeS = (orgS, orfS.id) if nodeQ in cbg.get_nodes() and nodeS in cbg.get_nodes(): pacbporf = cbg.get_pacbps_by_nodes(node1=nodeQ, node2=nodeS)[0] else: # take only the *best* HSP (highest scoring first one) hsp = alignment.hsps[0] # correct to absolute positions hsp.query_start = hsp.query_start + orfQ.protein_startPY hsp.sbjct_start = hsp.sbjct_start + orfS.protein_startPY # initialize the PacbP pacbporf = pacb.conversion.pacbp2pacbporf( pacb.PacbP(blastp_hsp=hsp), orfQ, orfS) ############################################################ if verbose: print "NEW:", pacbporf ############################################################ uqkey = pacbporf.construct_unique_key(nodeQ, nodeS) if not nodeQ in pacbpcol.get_nodes(): pacbpcol.add_node(nodeQ) if not nodeS in pacbpcol.get_nodes(): pacbpcol.add_node(nodeS) pacbpcol.add_edge(nodeQ, nodeS, wt=pacbporf.bitscore) # store to dpcpacbpcol -> pacbpcol is broken in pieces lateron! dpcpacbpcol.pacbps[(uqkey, nodeQ, nodeS)] = pacbporf # file cleanup _file_cleanup(multifastas.values()) _file_cleanup(["formatdb.log"]) _file_cleanup([fname + ".*" for fname in blastdbs.values()]) if not pacbpcol.organism_set_size() == cbg.organism_set_size(): ############################################################ if verbose: print "org_set_size() PCG < CBG" ############################################################ # no CBG on the reverse strand return None # ``deepcopy`` PacbPcollection dpcpacbpcol.add_nodes(pacbpcol.get_nodes()) for (uqkey, nodeQ, nodeS) in dpcpacbpcol.pacbps.keys(): (bitscore, length, orfQid, orfSid) = uqkey dpcpacbpcol.add_edge(nodeQ, nodeS, wt=bitscore) ############################################################################ if verbose: print pacbpcol, "bitscores:", print[pacbporf.bitscore for pacbporf in dpcpacbpcol.pacbps.values()] ############################################################################ # do some transformations on the pacbpcol pacbpcol.remove_low_connectivity_nodes(min_connectivity=cbg.node_count() - 1) splittedCBGs = pacbpcol.find_fully_connected_subgraphs( edges=cbg.node_count() - 1, max_missing_edges=0) # convert to list of CBGs and do some transformations cbgList = ListOfCodingBlockGraphs(splittedCBGs, input={}, crossdata={}) cbgList.remove_all_but_cbgs() cbgList.remove_cbgs_with_lt_nodes(cbg.node_count()) cbgList.harvest_pacbps_from_pacbpcollection(dpcpacbpcol) cbgList.remove_cbgs_without_omsr() cbgList.update_edge_weights_by_minimal_spanning_range() cbgList.order_graphlist_by_total_weight_and_identity() ############################################################################ if verbose: print "FScbgs (%s)" % len(cbgList) for fscbg in cbgList: print fscbg ############################################################################ if not cbgList: # no (better) frameshifted CBG return None elif cbgList and not cbgList[0].node_set().symmetric_difference( cbg.node_set()): # best CBG is not frameshifted, but CBG itself return None else: # score the difference between the frameshifted and current CBG score_cbg = cbg.total_weight() * cbg.omsr_identityscore() score_fscbg = cbgList[0].total_weight( ) * cbgList[0].omsr_identityscore() # check overlap between the frameshifted and current CBG a, b, c, d, e, f, g = relatively_positioned_towards(cbgList[0], cbg) ######################################################################## if verbose: print "CBG", cbg cbg.printmultiplealignment() for fscbg in cbgList: print "fsCBG:", fscbg fscbg.printmultiplealignment() ######################################################################## if (c, d) == ((0, 0, 1), (1, 0, 0)) or (c, d) == ((0, 0, 1), (1, 0, 0)): # CBG and frameshifted CBG do not share a single AA overlap... # This does not represent a frameshifted CBG as we searched for return False elif score_fscbg > score_cbg: # return the highest scoring, frameshifted CBG return cbgList[0] else: # no, still not convinced that this is a frameshifted CBG return False
def _blastorfset2blastdb(geneQ,geneS,blastdbfname,input,crossdata,GSgraph, blastoptions = None, elegiable_orfsQ=[], elegiable_orfsS_ids=[], logging=False): """ """ hitcnt = 0 for orfQ in elegiable_orfsQ: # check if protein sequence present in Orf object # in obscure cases of unigenes, no protein sequence present! if not orfQ.protein_sequence: continue # make unique node identifier and blast header nodeQ = (geneQ,orfQ.id) header = "%s_orf_%s" % (geneQ,orfQ.id) # do the blastp! blastrec = blastall_seq2db(header, orfQ.protein_sequence, dbname=blastdbfname, extra_blastp_params=blastoptions.extra_blastp_params ) # check if blast failed (then, blastrec == False) if not blastrec: continue # check if there are any hits/hsps! if len(blastrec.alignments) == 0: # no hits; continue continue for alignment in blastrec.alignments: # get back orfpointerB from the SBJCT and create nodeS _parts = alignment.title.split("_") geneS = "_".join(_parts[0:-2]).replace('>','') _orfpointerS = int(_parts[-1]) nodeS = (geneS,_orfpointerS) # ignore hit if nodeS orfid not occurring in the NON-empty list elegiable_orfsS_ids if elegiable_orfsS_ids and _orfpointerS not in elegiable_orfsS_ids: continue # get the Orf object of this sbjct sequence orfS = input[geneS]['orfs'].get_orf_by_id(_orfpointerS) # loop over the HSPs for hsp in alignment.hsps: # If hits are really tiny (happens in case of BLOSUM45 matrix), # discard them directly before precious time is lost... if len(hsp.query) <= blastoptions.BLASTP_DIRECTLY_IGNORE_TINY_HITS: continue # correct to absolute positions hsp.query_start = hsp.query_start + orfQ.protein_startPY hsp.sbjct_start = hsp.sbjct_start + orfS.protein_startPY hsp.query_end = hsp.query_end + orfQ.protein_startPY hsp.sbjct_end = hsp.sbjct_end + orfS.protein_startPY # VERY exceptional case: HSP starts or ends with a gap # I expect this is an error in Blastp .... strip_exterior_gaps(hsp) if hsp.query.find(" ") > 0: # VERY exceptional case: erroneously NCBI parsed HSP: # Score 8 (7 bits), expectation 1.7e+01, alignment length 41 # Query: 1622 STHTYDAC TRCI----PFVDTGHKHENPTEALLDSTA 1654 # TR P + H + P+++ S++ # Sbjct: 489 TEHIYLHT TRSTWPPKPPTNASHANTKPSKSHHRSSS 525 if len(hsp.query.split(" ")[-1]) == len(hsp.match): hsp.query = hsp.query.split(" ")[-1] hsp.sbjct = hsp.sbjct.split(" ")[-1] elif len(hsp.query.split(" ")[0]) == len(hsp.match): hsp.query = hsp.query.split(" ")[0] hsp.sbjct = hsp.sbjct.split(" ")[0] elif len(hsp.query) == len(hsp.match): # spaces in both query/match/sbjct while hsp.query.find(" ") > 0: pos = hsp.query.find(" ") if hsp.sbjct[pos] == " " and hsp.match[pos] == " ": hsp.query = hsp.query[0:pos] + hsp.query[pos+1:] hsp.match = hsp.match[0:pos] + hsp.match[pos+1:] hsp.sbjct = hsp.sbjct[0:pos] + hsp.sbjct[pos+1:] else: # HPS is not repairable -> quit trying break elif len(hsp.query) == len(hsp.sbjct): while hsp.query.find(" ") > 0: pos = hsp.query.find(" ") hsp.query = hsp.query[0:pos] + hsp.query[pos+1:] hsp.sbjct = hsp.sbjct[0:pos] + hsp.sbjct[pos+1:] # recreate alignment match string is done upon # creation of PacbP object else: pass # VERY exceptional case: HSP starts or ends with a gap # I expect this is an error in Blastp .... strip_exterior_gaps(hsp) try: pacbp = pacb.PacbP(blastp_hsp=hsp,MATRIX=blastoptions.MATRIX) except: # VERY exceptional miscelaneous cases: erroneously NCBI parsed HSP: print hsp print "'%s' X" % hsp.query, len(hsp.query), hsp.query_start, hsp.query_end print "'%s' X" % hsp.match, len(hsp.match) print "'%s' X" % hsp.sbjct, len(hsp.sbjct), hsp.sbjct_start, hsp.sbjct_end pacbp = pacb.PacbP(blastp_hsp=hsp,MATRIX=blastoptions.MATRIX) # make pacbp of this hsp pacbp = pacb.PacbP(blastp_hsp=hsp,MATRIX=blastoptions.MATRIX) # if logging is requested for, print this pacbp to STDOUT if logging: print ">>> Q", nodeQ, orfQ.tcode_symbolic(), "S", nodeS, orfS.tcode_symbolic(), pacbp, blastoptions.MATRIX.name, hsp.expect, hsp.bits print ">>>", blastoptions.extra_blastp_params if pacbp.length > 100: print pacbp.query[0:40]+'.'*7+str(pacbp.length-80)+'.'*7+pacbp.query[-40:] print pacbp.match[0:40]+'.'*7+str(pacbp.length-80)+'.'*7+pacbp.match[-40:] print pacbp.sbjct[0:40]+'.'*7+str(pacbp.length-80)+'.'*7+pacbp.sbjct[-40:] else: print pacbp.query print pacbp.match print pacbp.sbjct # blastoptions.BLASTP_HSP_MINIMAL_LENGTH represents the minimal # length of the aligned part. (To) short pacbp's are abandoned if pacbp.length < blastoptions.BLASTP_HSP_MINIMAL_LENGTH: if pacbp.identityscore == float(pacbp.length): # escape for 100% identical tiny pacbps pass elif pacbp.identity + pacbp.similarity == pacbp.length: # escape for 100% similar tiny pacbps pass else: # pacbp is to small. Discard! if logging: print "to small..." continue # check if the pacbp is not conflicting with the currect GSG graph # if so, ignore now because it will not yield a proper edge in an (accepted) CBG! if GSgraph and len(GSgraph) and GSgraph.is_pacbp_conflicting_with_genestructure(pacbp,orgQ=geneQ,orgS=geneS): ###print "GSGconflict!", nodeQ,nodeS, GSgraph.is_pacbp_conflicting_with_genestructure(pacbp,orgQ=geneQ,orgS=geneS), len(pacbp) continue # here we have a potentially accepted pacbp. # make a/the unique key of this pacbp key = (pacbp.bits, pacbp.length, orfQ.id,_orfpointerS) # check for evalue criterion if (blastoptions.BLASTP_HSP_MAXIMAL_EXPECT or blastoptions.BLASTP_HSP_MAXIMAL_EXPECT==0.0) and hsp.expect > blastoptions.BLASTP_HSP_MAXIMAL_EXPECT: # pacbp is long enough but has a to high evalue crossdata[(geneQ,geneS)]['lowscoring_pacbs'][key] = pacbp if logging: print "to low bitscore or expect" continue # check for bitscore criterion if (blastoptions.BLASTP_HSP_MINIMAL_BITS or blastoptions.BLASTP_HSP_MINIMAL_BITS==0) and pacbp.bits < blastoptions.BLASTP_HSP_MINIMAL_BITS: # pacbp is long enough but has a to low bitscore crossdata[(geneQ,geneS)]['lowscoring_pacbs'][key] = pacbp if logging: print "to low bitscore or expect" continue # !!Hurray!! an accepted pacbp. Store to crossdata # store it to the 'accepted_pacbs' dict of crossdata crossdata[(geneQ,geneS)]['accepted_pacbs'][key] = pacbp hitcnt+=1 if logging: print "ACCEPTED" # done -> check next orf! # return the filled crossdata structure return crossdata, hitcnt
def blastanalysescbgjunction( gsg, prevCBG, nextCBG, omit_cbg_orfs=False, omit_non_cbg_orfs=False, extra_blastp_params=CBG_JUNCTION_BLAST2PACBPCOL_EXTRA_BLASTP_PARAMS, omsr_2_mask_aa_length_correction=CBG_JUNCTION_BLAST2PACBPCOL_OMSR_2_AA_MASK, verbose=False): """ """ ############################################################ if verbose: stw = StopWatch('blastanalysescbgjunction') stw.start() ############################################################ orfs = {} if not omit_cbg_orfs: # gather Orfs from prevCBG and nextCBG for org, orflist, in prevCBG.get_orfs_of_graph().iteritems(): orf = orflist[0] orfs[(org, orf.id)] = orf for org, orflist, in nextCBG.get_orfs_of_graph().iteritems(): orf = orflist[0] orfs[(org, orf.id)] = orf ############################################################ if verbose: print stw.lap(), "orfs (1):", len(orfs) print _format_orf_nodes_to_string(orfs.keys()) ############################################################ # create masked fasta database in a dict fastadbmfa = parseFasta( create_hmmdb_for_neighbouring_cbgs( gsg.input, prevCBG, nextCBG, omsr_2_mask_aa_length_correction=omsr_2_mask_aa_length_correction, ).split("\n")) ############################################################ if verbose: print stw.lap(), "fasta db (1):", len(fastadbmfa) ############################################################ # remove ORFs that do not belong to prevCBG and nextCBG, # or that DO belong to prevCBG and nextCBG, or neither fastaheaders = fastadbmfa.keys() for header in fastaheaders: org, orfid = header.split("_orf_") orfid = int(orfid) node = (org, orfid) # check for the omit_non_cbg_orfs criterion add_orf = False if omit_non_cbg_orfs: if node not in orfs: del (fastadbmfa[header]) else: add_orf = True # check for the omit_cbg_orfs criterion if omit_cbg_orfs and node in orfs: del (fastadbmfa[header]) if add_orf: # get this Orf and add to orfs orfs[node] = gsg.input[org]['orfs'].get_orf_by_id(orfid) ############################################################ if verbose: print stw.lap(), "fasta db (2):", len(fastadbmfa) print _format_fastadbmfa_nodes_to_string(fastadbmfa.keys()) ############################################################ ############################################################ if verbose: print stw.lap(), "orfs (2):", len(orfs) print _format_orf_nodes_to_string(orfs.keys()) ############################################################ # no query/sbjct range left at all if not fastadbmfa: return [] # check if all organisms are still covered orgSet = Set([k.split("_orf_")[0] for k in fastadbmfa.keys()]) if orgSet.symmetric_difference(gsg.organism_set()): return [] # create !single! fasta database fastadbname = prevCBG.barcode() + "_" + nextCBG.barcode() + ".mfa" writeMultiFasta(fastadbmfa, fastadbname) formatdb(fname=fastadbname) # remap the identifiers of the orf objects i.o.t.... multifastas = {} blastdbs = {} pacbpcol = PacbpCollectionGraph() dpcpacbpcol = PacbpCollectionGraph() # ``deepcopied`` variant for pacbps ############################################################ if verbose: print stw.lap(), "blastp starting" ############################################################ for orgQ, orgS in prevCBG.pairwisecrosscombinations_organism(): for nodeQ, orfQ in orfs.iteritems(): # only blast the (masked) Orfs of orgQ if prevCBG.organism_by_node(nodeQ) != orgQ: continue # get the masked protein sequence of this orfObj header = orgQ + "_orf_" + str(orfQ.id) # check if key exists in fastadbmfa. In a case where # an Orf is masked out completely, it is absent here! if not fastadbmfa.has_key(header): continue protseq = fastadbmfa[orgQ + "_orf_" + str(orfQ.id)] # run blast_seqs2db blastrec = blastall_seq2db(orfQ.id, protseq, fastadbname, extra_blastp_params=extra_blastp_params) # omit empty blast records if len(blastrec.alignments) == 0: continue for alignment in blastrec.alignments: # get sbjct Org and Orf identifiers _orgS, _orfSid = alignment.title.replace(">", "").split("_orf_") if _orgS != orgS: continue nodeS = (_orgS, int(_orfSid)) orfS = orfs[nodeS] # take only the *best* HSP (highest scoring first one) hsp = alignment.hsps[0] # correct to absolute positions hsp.query_start = hsp.query_start + orfQ.protein_startPY hsp.sbjct_start = hsp.sbjct_start + orfS.protein_startPY # initialize the PacbP pacbporf = pacb.conversion.pacbp2pacbporf( pacb.PacbP(blastp_hsp=hsp), orfQ, orfS) ################################################################ if verbose: print pacbporf, orgQ, orgS, orfQ print pacbporf.query print pacbporf.match print pacbporf.sbjct ################################################################ # create nodes; ( Organism Identifier, Orf Identifier ) nodeQ = (orgQ, orfQ.id) nodeS = (orgS, orfS.id) uqkey = pacbporf.construct_unique_key(nodeQ, nodeS) if not nodeQ in pacbpcol.get_nodes(): pacbpcol.add_node(nodeQ) if not nodeS in pacbpcol.get_nodes(): pacbpcol.add_node(nodeS) pacbpcol.add_edge(nodeQ, nodeS, wt=pacbporf.bitscore) # store to dpcpacbpcol -> pacbpcol is broken in pieces lateron! dpcpacbpcol.pacbps[(uqkey, nodeQ, nodeS)] = pacbporf ############################################################ if verbose: print stw.lap(), "blastp done" ############################################################ # file cleanup _file_cleanup(multifastas.values()) _file_cleanup(["formatdb.log"]) _file_cleanup([fname + ".*" for fname in blastdbs.values()]) # check if all Organism/Gene identifiers are covered in PacbPs if not pacbpcol.organism_set_size() == gsg.organism_set_size(): return [] # ``deepcopy`` PacbPcollection pacbpcol to dpcpacbpcol # In dpcpacbpcol the actual PacbPORFs are stores & kept, # whereas pacbpcol itself is splitted in CBGs (which # function does not yet (!?) take the actual pacbps into account) dpcpacbpcol.add_nodes(pacbpcol.get_nodes()) for (uqkey, nodeQ, nodeS) in dpcpacbpcol.pacbps.keys(): (bitscore, length, orfQid, orfSid) = uqkey dpcpacbpcol.add_edge(nodeQ, nodeS, wt=bitscore) ################################################################ if verbose: print pacbpcol print "PCG bitscores:", print[p.bitscore for p in dpcpacbpcol.pacbps.values()] print "PCG nodes:", dpcpacbpcol.get_ordered_nodes() ################################################################ #### do some transformations on the pacbpcol ####pacbpcol.remove_low_connectivity_nodes(min_connectivity=gsg.EXACT_SG_NODE_COUNT-1) ####splittedCBGs = pacbpcol.find_fully_connected_subgraphs( #### edges=gsg.node_count()-1 , max_missing_edges=0 ) ##### convert to list of CBGs and do some transformations ####cbgList = ListOfCodingBlockGraphs(splittedCBGs,input={},crossdata={}) ####cbgList.remove_all_but_complete_cbgs() ####cbgList.remove_cbgs_with_lt_nodes(gsg.EXACT_SG_NODE_COUNT) ####cbgList.harvest_pacbps_from_pacbpcollection(dpcpacbpcol) ####cbgList.remove_cbgs_without_omsr() ####cbgList.update_edge_weights_by_minimal_spanning_range() ####cbgList.order_list_by_attribute(order_by='total_weight',reversed=True) min_connectivity = max([1, gsg.EXACT_SG_NODE_COUNT - 1 - 2]) pacbpcol.remove_low_connectivity_nodes(min_connectivity=min_connectivity) max_missing_edges = gsg.EXACT_SG_NODE_COUNT - 3 splittedCBGs = pacbpcol.find_fully_connected_subgraphs( edges=gsg.node_count() - 1, max_missing_edges=max_missing_edges) # convert to list of CBGs and do some transformations cbgList = ListOfCodingBlockGraphs(splittedCBGs, input={}, crossdata={}) cbgList.remove_all_but_cbgs() cbgList.harvest_pacbps_from_pacbpcollection(dpcpacbpcol) cbgList.make_pacbps_for_missing_edges() cbgList.remove_all_but_complete_cbgs() cbgList.remove_cbgs_with_lt_nodes(gsg.EXACT_SG_NODE_COUNT) cbgList.remove_cbgs_without_omsr() cbgList.update_edge_weights_by_minimal_spanning_range() cbgList.order_list_by_attribute(order_by='total_weight', reversed=True) # and create_cache() for these CBGs for cbg in cbgList: cbg.create_cache() #################################################################### if verbose: print stw.lap(), "CBGs created", len(cbgList) for newcbg in cbgList: print "new:", newcbg #################################################################### # return list with CBGs return cbgList.codingblockgraphs
def get_reverse_cbg(cbg, frame, verbose=False): """ Get the ReversecomplementCodingBlockGraph in requested frame of this CBG @type cbg: CodingBlockGraph @param cbg: CodingBlockGraph to reversecomplement @type frame: integer @param frame: 0,1 or 2 @type verbose: Boolean @param verbose: print intermediate info to STDOUT for debugging purposes @rtype: ReversecomplementCodingBlockGraph or None @return: ReversecomplementCodingBlockGraph (when existing) or None """ min_orf_length = (cbg.omsrlength() / 2) * 3 orfs = get_reverse_strand_orfsets(cbg, frame, min_orf_length=min_orf_length) # remap the identifiers of the orf objects i.o.t.... multifastas = {} blastdbs = {} pacbpcol = PacbpCollectionGraph() dpcpacbpcol = PacbpCollectionGraph() # ``deepcopied`` variant for pacbps for org in orfs.keys(): fname = "%s_reversecbg_%s.mfa" % (org, cbg.barcode()) writeMultiFasta(orfs[org].tofastadict(), fname) multifastas[org] = fname ######################################################################## if verbose: print "ORFS:", org, len(orfs[org].orfs), print[len(o.protein_sequence) for o in orfs[org].orfs] ######################################################################## revpacbps = {} for orgQ, orgS in cbg.pairwisecrosscombinations_organism(): # create blastdb if it does not exist yet if not blastdbs.has_key(orgS): formatdb(fname=multifastas[orgS]) blastdbs[orgS] = multifastas[orgS] revpacbporfs = {} for orfQ in orfs[orgQ].orfs: # run blast_seqs2db blastrec = blastall_seq2db(orfQ.id, orfQ.protein_sequence, dbname="./" + blastdbs[orgS]) if len(blastrec.alignments) == 0: continue for alignment in blastrec.alignments: # obtain coordinates from sbjct orf identifier orfS = orfs[orgS].get_orf_by_id( alignment.title.replace(">", "")) # take only the *best* HSP (highest scoring first one) hsp = alignment.hsps[0] # skip if hsp is very short if len(hsp.query) < cbg.omsrlength() / 2: continue # correct to absolute positions hsp.query_start = hsp.query_start + orfQ.protein_startPY hsp.sbjct_start = hsp.sbjct_start + orfS.protein_startPY # initialize the PacbP pacbporf = pacb.conversion.pacbp2pacbporf( pacb.PacbP(blastp_hsp=hsp), orfQ, orfS) ################################################################ if verbose: print pacbporf, orgQ, orgS, orfQ print pacbporf.query print pacbporf.match print pacbporf.sbjct ###pacbporf.print_protein_and_dna() ################################################################ nodeQ = (orgQ, orfQ.protein_startPY) nodeS = (orgS, orfS.protein_startPY) uqkey = pacbporf.construct_unique_key(nodeQ, nodeS) if not nodeQ in pacbpcol.get_nodes(): pacbpcol.add_node(nodeQ) if not nodeS in pacbpcol.get_nodes(): pacbpcol.add_node(nodeS) pacbpcol.add_edge(nodeQ, nodeS, wt=pacbporf.bitscore) # store to dpcpacbpcol -> pacbpcol is broken in pieces lateron! dpcpacbpcol.pacbps[(uqkey, nodeQ, nodeS)] = pacbporf # file cleanup _file_cleanup(multifastas.values()) _file_cleanup(["formatdb.log"]) _file_cleanup([fname + ".*" for fname in blastdbs.values()]) if not pacbpcol.organism_set_size() == cbg.organism_set_size(): # no CBG on the reverse strand return None # ``deepcopy`` PacbPcollection dpcpacbpcol.add_nodes(pacbpcol.get_nodes()) for (uqkey, nodeQ, nodeS) in dpcpacbpcol.pacbps.keys(): (bitscore, length, orfQid, orfSid) = uqkey dpcpacbpcol.add_edge(nodeQ, nodeS, wt=bitscore) ############################################################################ if verbose: print pacbpcol, "bitscores:", print[pacbporf.bitscore for pacbporf in dpcpacbpcol.pacbps.values()] ############################################################################ # do some transformations on the pacbpcol pacbpcol.remove_low_connectivity_nodes(min_connectivity=cbg.node_count() - 1) splittedCBGs = pacbpcol.find_fully_connected_subgraphs( edges=cbg.node_count() - 1, max_missing_edges=0) # convert to list of CBGs and do some transformations cbgList = ListOfCodingBlockGraphs(splittedCBGs, input={}, crossdata={}) cbgList.remove_all_but_complete_cbgs() cbgList.harvest_pacbps_from_pacbpcollection(dpcpacbpcol) cbgList.remove_cbgs_without_omsr() cbgList.update_edge_weights_by_minimal_spanning_range() cbgList.order_list_by_attribute(order_by='total_weight', reversed=True) ############################################################################ if verbose: for revcbg in cbgList: print "revCBG:", revcbg ############################################################################ if not cbgList: # no CBG on the reverse strand return None else: # return the highest scoring CBG as a ReversecomlementCodingBlockGraph return CodingBlockGraph2ReversecomlementCodingBlockGraph( cbgList.codingblockgraphs[0])
######################################################################## if verbose: print "ORFS:", org, len(orfs[org].orfs), print [ orf.id for orf in orfs[org].orfs ], print [ str(orf) for orf in orfs[org].orfs ] ######################################################################## for orgQ,orgS in cbg.pairwisecrosscombinations_organism(): # create blastdb if it does not exist yet if not blastdbs.has_key(orgS): formatdb(fname=multifastas[orgS]) blastdbs[orgS] = multifastas[orgS] for orfQ in orfs[orgQ].orfs: # run blast_seqs2db blastrec = blastall_seq2db(orfQ.id,orfQ.protein_sequence, dbname="./"+blastdbs[orgS]) if len(blastrec.alignments) == 0: continue for alignment in blastrec.alignments: # obtain coordinates from sbjct orf identifier orfid = alignment.title.replace(">","").split(" ")[0].replace("_","") orfS = orfs[orgS].get_orf_by_id(int(orfid)) nodeQ = ( orgQ, orfQ.id ) nodeS = ( orgS, orfS.id ) if nodeQ in cbg.get_nodes() and nodeS in cbg.get_nodes(): pacbporf = cbg.get_pacbps_by_nodes(node1=nodeQ,node2=nodeS)[0] else: # take only the *best* HSP (highest scoring first one) hsp = alignment.hsps[0]
def blastanalysescbgjunction(gsg,prevCBG,nextCBG, omit_cbg_orfs = False, omit_non_cbg_orfs = False, extra_blastp_params=CBG_JUNCTION_BLAST2PACBPCOL_EXTRA_BLASTP_PARAMS, omsr_2_mask_aa_length_correction=CBG_JUNCTION_BLAST2PACBPCOL_OMSR_2_AA_MASK, verbose=False): """ """ ############################################################ if verbose: stw = StopWatch('blastanalysescbgjunction') stw.start() ############################################################ orfs = {} if not omit_cbg_orfs: # gather Orfs from prevCBG and nextCBG for org,orflist, in prevCBG.get_orfs_of_graph().iteritems(): orf = orflist[0] orfs[(org,orf.id)] = orf for org,orflist, in nextCBG.get_orfs_of_graph().iteritems(): orf = orflist[0] orfs[(org,orf.id)] = orf ############################################################ if verbose: print stw.lap(), "orfs (1):",len(orfs) print _format_orf_nodes_to_string(orfs.keys()) ############################################################ # create masked fasta database in a dict fastadbmfa = parseFasta( create_hmmdb_for_neighbouring_cbgs( gsg.input,prevCBG,nextCBG, omsr_2_mask_aa_length_correction=omsr_2_mask_aa_length_correction, ).split("\n") ) ############################################################ if verbose: print stw.lap(), "fasta db (1):",len(fastadbmfa) ############################################################ # remove ORFs that do not belong to prevCBG and nextCBG, # or that DO belong to prevCBG and nextCBG, or neither fastaheaders = fastadbmfa.keys() for header in fastaheaders: org,orfid = header.split("_orf_") orfid = int(orfid) node = (org,orfid) # check for the omit_non_cbg_orfs criterion add_orf = False if omit_non_cbg_orfs: if node not in orfs: del(fastadbmfa[header]) else: add_orf = True # check for the omit_cbg_orfs criterion if omit_cbg_orfs and node in orfs: del(fastadbmfa[header]) if add_orf: # get this Orf and add to orfs orfs[node] = gsg.input[org]['orfs'].get_orf_by_id(orfid) ############################################################ if verbose: print stw.lap(), "fasta db (2):",len(fastadbmfa) print _format_fastadbmfa_nodes_to_string(fastadbmfa.keys()) ############################################################ ############################################################ if verbose: print stw.lap(), "orfs (2):",len(orfs) print _format_orf_nodes_to_string(orfs.keys()) ############################################################ # no query/sbjct range left at all if not fastadbmfa: return [] # check if all organisms are still covered orgSet = Set([ k.split("_orf_")[0] for k in fastadbmfa.keys()]) if orgSet.symmetric_difference(gsg.organism_set()): return [] # create !single! fasta database fastadbname = prevCBG.barcode()+"_"+nextCBG.barcode()+".mfa" writeMultiFasta(fastadbmfa,fastadbname) formatdb(fname=fastadbname) # remap the identifiers of the orf objects i.o.t.... multifastas = {} blastdbs = {} pacbpcol = PacbpCollectionGraph() dpcpacbpcol = PacbpCollectionGraph() # ``deepcopied`` variant for pacbps ############################################################ if verbose: print stw.lap(), "blastp starting" ############################################################ for orgQ,orgS in prevCBG.pairwisecrosscombinations_organism(): for nodeQ,orfQ in orfs.iteritems(): # only blast the (masked) Orfs of orgQ if prevCBG.organism_by_node(nodeQ) != orgQ: continue # get the masked protein sequence of this orfObj header = orgQ+"_orf_"+str(orfQ.id) # check if key exists in fastadbmfa. In a case where # an Orf is masked out completely, it is absent here! if not fastadbmfa.has_key(header): continue protseq = fastadbmfa[orgQ+"_orf_"+str(orfQ.id)] # run blast_seqs2db blastrec = blastall_seq2db(orfQ.id,protseq,fastadbname, extra_blastp_params=extra_blastp_params) # omit empty blast records if len(blastrec.alignments) == 0: continue for alignment in blastrec.alignments: # get sbjct Org and Orf identifiers _orgS,_orfSid = alignment.title.replace(">","").split("_orf_") if _orgS != orgS: continue nodeS = (_orgS,int(_orfSid)) orfS = orfs[nodeS] # take only the *best* HSP (highest scoring first one) hsp = alignment.hsps[0] # correct to absolute positions hsp.query_start = hsp.query_start + orfQ.protein_startPY hsp.sbjct_start = hsp.sbjct_start + orfS.protein_startPY # initialize the PacbP pacbporf = pacb.conversion.pacbp2pacbporf( pacb.PacbP(blastp_hsp=hsp),orfQ,orfS) ################################################################ if verbose: print pacbporf, orgQ,orgS, orfQ print pacbporf.query print pacbporf.match print pacbporf.sbjct ################################################################ # create nodes; ( Organism Identifier, Orf Identifier ) nodeQ = ( orgQ, orfQ.id ) nodeS = ( orgS, orfS.id ) uqkey = pacbporf.construct_unique_key(nodeQ,nodeS) if not nodeQ in pacbpcol.get_nodes(): pacbpcol.add_node(nodeQ) if not nodeS in pacbpcol.get_nodes(): pacbpcol.add_node(nodeS) pacbpcol.add_edge(nodeQ,nodeS,wt=pacbporf.bitscore) # store to dpcpacbpcol -> pacbpcol is broken in pieces lateron! dpcpacbpcol.pacbps[(uqkey,nodeQ,nodeS)] = pacbporf ############################################################ if verbose: print stw.lap(), "blastp done" ############################################################ # file cleanup _file_cleanup(multifastas.values()) _file_cleanup(["formatdb.log"]) _file_cleanup([ fname+".*" for fname in blastdbs.values()]) # check if all Organism/Gene identifiers are covered in PacbPs if not pacbpcol.organism_set_size() == gsg.organism_set_size(): return [] # ``deepcopy`` PacbPcollection pacbpcol to dpcpacbpcol # In dpcpacbpcol the actual PacbPORFs are stores & kept, # whereas pacbpcol itself is splitted in CBGs (which # function does not yet (!?) take the actual pacbps into account) dpcpacbpcol.add_nodes( pacbpcol.get_nodes() ) for (uqkey,nodeQ,nodeS) in dpcpacbpcol.pacbps.keys(): (bitscore,length,orfQid,orfSid) = uqkey dpcpacbpcol.add_edge(nodeQ,nodeS,wt=bitscore) ################################################################ if verbose: print pacbpcol print "PCG bitscores:", print [ p.bitscore for p in dpcpacbpcol.pacbps.values() ]