from Graph_gff import Segments, Features, write_line from Functions import * ### functions to generate a genome's gff from the graph's gff # functions to get the detailed gff with all the fragments of the features # outputs each fragment of the feature in a gff def gff_detail(list_seg,feature_id): # loop that goes through all the segments that have the current feature # keeps the first segment present in the genome found, and when it finds a segment absent in the genome, prints the current part of the fragment, and resets the first segment present. # continues to go through the segments, keeps the next segment present in the genome, and when it finds a segment absent, prints the second part of the feature, etc. # at the end of the loop, prints the last part of the fragment. [feat_start,seg_start,last_seg]=["","",""] # first segment of the feature, first segment of the current part of the feature, last segment in the part of the feature, for loop below for segment in list_seg: if segment[1:] in segments_on_target_genome: if feat_start=="": feat_start=segment[1:] if seg_start=="": # if we dont have a start, take the current segment for the start of the part of the feature seg_start=segment[1:] #else: if we already have a start, keep going though the segments until we find a stop (segment not in azucena) else: if seg_start!="": # found a stop and we have a start. print the line, reset seg_start, and keep going through the segments to find the next seg_start out_line=create_line_detail(last_seg,feature_id,seg_start,feat_start) write_line(out_line,output_detail_gff,False) seg_start="" #else: if the current segment is not in azucena but there is no start, keep looking for a start last_seg=segment if last_seg[1:] in segments_on_target_genome: out_line=create_line_detail(list_seg[-1],feature_id,seg_start,feat_start) write_line(out_line,output_detail_gff,False) # functions to get the gff with one line per feature # outputs the feature once in a gff, from the first to the last segment present on the new genome (if the size is ok) : def gff_one(first_seg,last_seg,feature_id,list_seg,max_diff): if (first_seg!=''): # feature present on the target genome size_on_new_genome=get_feature_stop_on_genome(last_seg,feature_id)-get_feature_start_on_genome(first_seg,feature_id)+1 size_diff=abs(size_on_new_genome-Features[feature_id].size) if right_size(size_on_new_genome,max_diff,feature_id): line=create_line_gff_one(first_seg,last_seg,feature_id,size_diff,list_seg) write_line(line,output_gff_one,False) return size_diff else: return "bad_size" else : return "absent" # writes the gff of azucena using the gff of the graph def genome_gff(pos_seg, gff, gfa, out_once, out_detail, out_var,target_genome_name): print("generation of the genome's gff ") # create variables and open files [once,detail,var,stats]=[True,True,True,False] if var: [paths,seg_seq]=get_segments_sequence_and_paths(gfa) file_out_var = open(out_var, 'w') global output_variations output_variations=[0,"",file_out_var] if detail: file_out_detail = open(out_detail, 'w') global output_detail_gff output_detail_gff=[0,"",file_out_detail] if once: file_out=open(out_once,'w') global output_gff_one output_gff_one=[0,"",file_out] bad_size_features=0 absent_features=0 diff_size_transfered_features=[0,0] # [count,sum], to get the average if stats: # create objects for stats on how many segments are absent in azucena, their average length, etc feature_missing_segments=[[],[],[],[],[],[],[]] # [feature_missing_first,feature_missing_middle,feature_missing_last,feature_missing_all,feature_missing_total,feature_total,feature_ok] # the fist segment of the feature is missing - feature_missing_first # the last segment of the feature is missing - feature_missing_last # at least one middle segment of the feature is missing - feature_missing_middle # the entire feature is missing - feature_missing_all # at least one segment is missing first, last, or middle) - feature_missing_total # no segment is missing, the feature is complete - feature_ok # total number of features, with missing segments or not - feature_total get_segments_positions_on_genome(pos_seg) list_feature_to_transfer=get_all_features_in_gff(gff) # get the list of all the features to transfer from the gff for feat in list_feature_to_transfer: # for each feature, get list of the segments where it is and the first and last segment of the feature on the new genome list_seg=Features[feat].segments_list first_seg=get_first_seg(list_seg) last_seg=get_first_seg(reversed(list_seg)) # outputs the feature once in a gff, from the first to the last segment present on the new genome if once: max_diff=2 # maximum difference size (n times bigger of smaller) transfer_stat=gff_one(first_seg,last_seg,feat,list_seg,max_diff) # insertion not considered !!! if transfer_stat=="bad_size": bad_size_features+=1 elif transfer_stat=="absent": absent_features+=1 else: diff_size_transfered_features[0]+=1 diff_size_transfered_features[1]+=transfer_stat write_line("",output_gff_one,True) # outputs each fragment of the feature in a gff if detail: gff_detail(list_seg,feat) # insertions ! write_line("",output_detail_gff,True) # outputs the detail of variations of the feature : if var: print_variations(first_seg,last_seg,feat,paths,seg_seq,target_genome_name) write_line("",output_variations,True) if stats==True: stats_feature_missing_segment(feature_missing_segments,first_seg,last_seg,list_seg,feat) # close output_files if detail: file_out_detail.close() if once: file_out.close() if var: file_out_var.close() # print stats if stats==True: if once: print(len(Features)-(bad_size_features+absent_features),"out of",len(Features),"features are transfered.") print(bad_size_features,"out of",len(Features), "features are not transfered because they are too big or too small compared to the original genome.") print(absent_features,"out of",len(Features),"features are not transfered because they are absent in the new genome.") print("average length difference of the transfered genes : ",diff_size_transfered_features[1]/diff_size_transfered_features[0]) stats_features(feature_missing_segments) # functions to get the detail of the variations in the features def print_variations(first_seg,last_seg,feat,paths,seg_seq,target_genome_name): if (first_seg!=''): # if the feature is not completly absent # add the else, output absent features [variation,list_segfeat_nb,list_segfeat_azu]=create_var(feat,first_seg,last_seg,paths,target_genome_name) # removes the strands in the segment lists feature=Features[feat] feat_start=feature.start # loop to go through both paths with i and j [i,j,var_count]=[0,0,0] # detect and print variations ignoring the strands while (i<len(list_segfeat_nb)) & (j<len(list_segfeat_azu)): if list_segfeat_nb[i] != list_segfeat_azu[j]: # if there is a difference between the two paths if list_segfeat_azu[j] not in list_segfeat_nb: # if the segment in azu is absent in nb if list_segfeat_nb[i] not in list_segfeat_azu: # if the segment in nb is absent is azu # if both segments are absent in the other genome, its a substitution variation.last_seg_in_target=list_segfeat_azu[j][1:] if (variation.type=='insertion') | (variation.type=='deletion'): # print the current variation before treating the substitution print_current_var(variation,feat_start,list_segfeat_azu,feat,i) reset_var(variation) var_count+=1 if variation.type=='substitution': continue_var(variation,seg_seq,list_segfeat_nb,list_segfeat_azu,i,j,0) else: # initiate substitution init_new_var(variation,"substitution",list_segfeat_nb,list_segfeat_azu,i,j,seg_seq,feature) i+=1;j+=1 else: # azu segment not in nb, but nb segment in azu : insertion or continue substitution if variation.type=='deletion': # print the current variation before treating the insertion print_current_var(variation,feat_start,list_segfeat_azu,feat,i) reset_var(variation) var_count+=1 variation.last_seg_in_target=list_segfeat_azu[j][1:] if variation.type=='insertion': continue_var(variation,seg_seq,list_segfeat_nb,list_segfeat_azu,i,j,0) elif variation.type=="substitution": while list_segfeat_azu[j]!=list_segfeat_nb[i]: continue_var(variation,seg_seq,list_segfeat_nb,list_segfeat_azu,i,j,2) j+=1 print_current_var(variation,feat_start,list_segfeat_azu,feat,i) reset_var(variation) var_count+=1 j-=1 else: # intiate insertion init_new_var(variation,"insertion",list_segfeat_nb,list_segfeat_azu,i,j,seg_seq,feature) j+=1 elif list_segfeat_nb[i] not in list_segfeat_azu: # nb segment not in azu, but azu segment in nb : deletion or continue substitution if variation.type=='insertion': # print the current variation before treating the deletion print_current_var(variation,feat_start,list_segfeat_azu,feat,i) reset_var(variation) var_count+=1 if variation.type=='deletion': continue_var(variation,seg_seq,list_segfeat_nb,list_segfeat_azu,i,j,0) elif variation.type=="substitution": while list_segfeat_azu[j]!=list_segfeat_nb[i]: continue_var(variation,seg_seq,list_segfeat_nb,list_segfeat_azu,i,j,1) i+=1 print_current_var(variation,feat_start,list_segfeat_azu,feat,i) reset_var(variation) var_count+=1 i-=1 else: # intiate deletion init_new_var(variation,"deletion",list_segfeat_nb,list_segfeat_azu,i,j,seg_seq,feature) i+=1 else : # if both segments are present in the other genome but not at the same position. weird case never found yet # can be a substitution. check later if its not an inversion variation.last_seg_in_target=list_segfeat_azu[j][1:] if (variation.type=='insertion') | (variation.type=='deletion'): # print the current variation before treating the substitution print_current_var(variation,feat_start,list_segfeat_azu,feat,i) reset_var(variation) var_count+=1 if variation.type=='substitution': continue_var(variation,seg_seq,list_segfeat_nb,list_segfeat_azu,i,j,0) else: # initiate substitution init_new_var(variation,"substitution",list_segfeat_nb,list_segfeat_azu,i,j,seg_seq,feature) i+=1;j+=1 else: # segment present in both, no variation. print the running indel if there is one if variation.type!='': # print the current variation if there is one print_current_var(variation,feat_start,list_segfeat_azu,feat,i) var_count+=1 reset_var(variation) variation.last_seg_in_target=list_segfeat_azu[j][1:] i+=1;j+=1 if (variation.type!=''): # if there was a current variation when we reached the end, print it print_current_var(variation,feat_start,list_segfeat_azu,feat,i) var_count+=1 reset_var(variation) if i<=len(list_segfeat_nb)-1: # if we didn't reach the length of the segment list for the first genome, the end is missing for the second genome print_last_deletion(variation,list_segfeat_nb,i,feat_start,feature,seg_seq) var_count+=1 reset_var(variation) if var_count==0: # if no variation was encountered print_novar(variation) def print_current_var(variation,feat_start,list_segfeat_azu,feat,i): warning=detect_small_inversion(variation) if variation.type=='insertion': [pos_old,pos_new]=get_old_new_pos_insertion(variation,feat_start,list_segfeat_azu,feat) line=f'{variation.feature_id}\t{variation.feature_type}\t{variation.chr}\t{variation.start_new}\t{variation.stop_new}\t{variation.size_new}\t{variation.inversion}\t{variation.size_diff}\tinsertion\t-\t{variation.alt}\t{len(variation.alt)}\t{pos_old}\t{pos_new}{warning}\n' write_line(line,output_variations,False) elif variation.type=='deletion': [pos_old,pos_new]=get_old_new_pos_deletion(variation,feat_start,list_segfeat_azu,feat) line=f'{variation.feature_id}\t{variation.feature_type}\t{variation.chr}\t{variation.start_new}\t{variation.stop_new}\t{variation.size_new}\t{variation.inversion}\t{variation.size_diff}\tdeletion\t{variation.ref}\t-\t{len(variation.ref)}\t{pos_old}\t{pos_new}{warning}\n' write_line(line,output_variations,False) elif variation.type=='substitution': [pos_old,pos_new]=get_old_new_pos_substitution(feat_start,variation,list_segfeat_azu,feat) size_subs=f'{len(variation.ref)}/{len(variation.alt)}' line=f'{variation.feature_id}\t{variation.feature_type}\t{variation.chr}\t{variation.start_new}\t{variation.stop_new}\t{variation.size_new}\t{variation.inversion}\t{variation.size_diff}\tsubstitution\t{variation.ref}\t{variation.alt}\t{size_subs}\t{pos_old}\t{pos_new}{warning}\n' # print the substitutions of different size as deletion+insertion. #if len(variation.ref) == len(variation.alt): # if the substituion is between two segment of the same size, print it # size_subs=len(variation.ref) # line=f'{variation.feature_id}\t{variation.feature_type}\t{variation.chr}\t{variation.start_new}\t{variation.stop_new}\t{variation.size_new}\t{variation.inversion}\t{variation.size_diff}\tsubstitution\t{variation.ref}\t{variation.alt}\t{size_subs}\t{pos_old}\t{pos_new}{warning}\n' #else : # # if the segments of the substitution have a different size, print deletion then insertion at the same position. # line=f'{variation.feature_id}\t{variation.feature_type}\t{variation.chr}\t{variation.start_new}\t{variation.stop_new}\t{variation.size_new}\t{variation.inversion}\t{variation.size_diff}\tdeletion\t{variation.ref}\t-\t{len(variation.ref)}\t{pos_old}\t{pos_new}{warning}\n' # line+=f'{variation.feature_id}\t{variation.feature_type}\t{variation.chr}\t{variation.start_new}\t{variation.stop_new}\t{variation.size_new}\t{variation.inversion}\t{variation.size_diff}\tinsertion\t-\t{variation.alt}\t{len(variation.alt)}\t{pos_old}\t{pos_new}{warning}\n' write_line(line,output_variations,False) def detect_small_inversion(variation): [list_ref_common,list_alt_common]=[list(),list()] list_ref_unstrand=[segment_stranded[1:] for segment_stranded in variation.seg_ref] list_alt_unstrand=[segment_stranded[1:] for segment_stranded in variation.seg_alt] for seg in variation.seg_ref: if seg[1:] in list_alt_unstrand: list_ref_common.append(seg) for seg in variation.seg_alt: if seg[1:] in list_ref_unstrand: list_alt_common.append(seg) if (len(list_ref_common)>len(list_ref_unstrand)*0.5) & (len(list_alt_common)>len(list_alt_unstrand)*0.5): return f'\t# Suspected inversion within this substitution.' else: return '' def print_last_deletion(variation,list_segfeat_nb,i,feat_start,feature,seg_seq): pos_old=int(Segments[list_segfeat_nb[i][1:]].start)-int(feat_start)+1 del_sequence=get_sequence_list_seg(list_segfeat_nb,i,feature,seg_seq) length=len(del_sequence) pos_new=str(int(variation.size_new)+1) # the deletion is at the end of the feature on the new genome line=f'{variation.feature_id}\t{variation.feature_type}\t{variation.chr}\t{variation.start_new}\t{variation.stop_new}\t{variation.size_new}\t{variation.inversion}\t{variation.size_diff}\tdeletion\t{del_sequence}\t-\t{length}\t{pos_old}\t{pos_new}\n' write_line(line,output_variations,False) def print_novar(variation): line=f'{variation.feature_id}\t{variation.feature_type}\t{variation.chr}\t{variation.start_new}\t{variation.stop_new}\t{variation.size_new}\t{variation.inversion}\t{variation.size_diff}\tno_var\t-\t-\t-\t-\t-\n' write_line(line,output_variations,False) # not used. def get_list_segments_missing(list_seg,segments_on_target_genome): segments_missing=[] for segment in list_seg: if segment[1:] not in segments_on_target_genome: segments_missing.append(Segments[segment[1:]]) return segments_missing # takes a feature and a feature type, returns a list of child features that have the wanted type. currently not used. def get_child_list(feature,child_type): if type=="": return feature.childs list_childs=[] for child in feature.childs: if Features[child].type==child_type: list_childs.append(child) return list_childs