def create_illumina_genotyping_references(illumina_read_dir='../Genotyping/'):
from reference_vntr import load_unique_vntrs_data
reference_vntrs = load_unique_vntrs_data()
id_to_gene = {1220: 'GP1BA', 1221: 'CSTB', 1214: 'MAOA'}
repeats = {'GP1BA': range(1, 5), 'CSTB': range(1, 16), 'MAOA': range(1, 6)}
repeats_patterns = {
'GP1BA': [
'AGCCCGACCACCCCAGAGCCCACCTCAGAGCCCGCCCCC',
'AGCCCGACCACCCCGGAGCCCACCTCAGAGCCCGCCCCC',
'AGCCCGACCACCCCGGAGCCCACCCCAATCCCGACCATCGCCA'
],
'CSTB':
['CGCGGGGCGGGG', 'CGCGGGGCGGGG', 'CGCGGGGCGGGG', 'CGGCGGGCGGGG'],
'MAOA': [
'ACCGGCACCGGCACCAGTACCCGCACCAGT', 'ACCGGCACCGGCACCGAGCGCAAGGCGGAG',
'ACCGGCACCGGCACCAGTACCCGCACCAGT'
]
}
for vntr_id in id_to_gene.keys():
# if vntr_id != 1221:
# continue
for repeat in repeats[id_to_gene[vntr_id]]:
outfile = illumina_read_dir + id_to_gene[vntr_id] + '/' + str(
repeat) + '.pacfa'
create_reference_region_with_specific_repeats(
reference_vntrs[vntr_id], repeat, outfile, 3000,
repeats_patterns[id_to_gene[vntr_id]])
def create_illumina_copy_number_variation_references(
illumina_read_dir='../Illumina_copy_number/'):
from reference_vntr import load_unique_vntrs_data
reference_vntrs = load_unique_vntrs_data()
id_to_gene = {
119: 'DRD4',
1220: 'GP1BA',
1221: 'CSTB',
1214: 'MAOA',
1219: 'IL1RN'
}
repeats = {
'DRD4': range(1, 12),
'GP1BA': range(1, 6),
'CSTB': range(1, 16),
'MAOA': range(1, 6),
'IL1RN': range(1, 10)
}
for vntr_id in id_to_gene.keys():
for repeat in repeats[id_to_gene[vntr_id]]:
outfile = illumina_read_dir + id_to_gene[vntr_id] + '/' + str(
repeat) + '.fa'
create_reference_region_with_specific_repeats(
reference_vntrs[vntr_id], repeat, outfile, 149)
def check_trio_consistency(father_file, mother_file, child_file):
father_genotypes = get_genotypes(father_file)
mother_genotypes = get_genotypes(mother_file)
child_genotypes = get_genotypes(child_file)
from reference_vntr import load_unique_vntrs_data
reference_vntrs = load_unique_vntrs_data()
vntr_ids = set(father_genotypes.keys() + mother_genotypes.keys() +
child_genotypes.keys())
print('Total vntrs: %s' % len(vntr_ids))
inconsistents = []
consistents = []
for vid in vntr_ids:
if not is_consistent(vid, father_genotypes, mother_genotypes,
child_genotypes):
inconsistents.append(
len(reference_vntrs[vid].get_repeat_segments()))
# print (len(reference_vntrs[vid].pattern))
if 83 <= len(reference_vntrs[vid].pattern) < 85:
print reference_vntrs[vid].left_flanking_region[-50:]
# print('%s: %s %s %s' % (vid, father_genotypes[vid], mother_genotypes[vid], child_genotypes[vid]))
else:
consistents.append(
len((reference_vntrs[vid].get_repeat_segments())))
print('Total inconsistencies: %s' % len(inconsistents))
print(consistents)
print(inconsistents)
def get_pacbio_comparison_result():
reference_vntrs = load_unique_vntrs_data()
id_to_gene = {1221: 'CSTB', 1216: 'HIC1', 1215: 'INS'}
genes = glob.glob('../Pacbio_copy_number/*')
for gene_dir in genes:
print(gene_dir)
files = glob.glob(gene_dir + '/*30x.fastq.sam')
gene_name = gene_dir.split('/')[-1]
mapped_reads = {}
for file_name in files:
copies = int(file_name.split('_')[-2])
make_bam_and_index(file_name)
base_name = file_name[:-4]
original_bam = base_name + '.bam'
bwasw_alignment(base_name)
blasr_alignment(base_name)
bwasw_alignment_file = base_name[:-3] + '_bwasw_aln.bam'
blasr_alignment_file = base_name[:-3] + '_blasr_aln.bam'
vntr_id = None
for vid, gname in id_to_gene.items():
if gname == gene_name:
vntr_id = vid
ref_length = copies * len(reference_vntrs[vntr_id].pattern) + 2000
true_ids = get_pacbio_true_read_ids(original_bam,
reference_vntrs[vntr_id],
ref_length)
blasr_ids = get_id_of_reads_mapped_to_vntr_in_bamfile(
blasr_alignment_file, reference_vntrs[vntr_id])
bwasw_ids = get_id_of_reads_mapped_to_vntr_in_bamfile(
bwasw_alignment_file, reference_vntrs[vntr_id])
blasr_tp = [
read_id for read_id in blasr_ids if read_id in true_ids
]
bwasw_tp = [
read_id for read_id in bwasw_ids if read_id in true_ids
]
vntr_finder = VNTRFinder(reference_vntrs[vntr_id])
our_filtering = get_out_pacbio_filtered_counts(
base_name, vntr_finder)
our_selection = our_filtering
mapped_reads[copies] = [
len(true_ids), our_filtering, our_selection,
len(bwasw_tp),
len(blasr_tp)
]
with open(gene_dir + '/result.txt', 'w') as out:
for copies in sorted(mapped_reads.iterkeys()):
original, our_filtering, our_selection, bwasw, blasr = mapped_reads[
copies]
out.write('%s %s %s %s %s %s\n' %
(copies, original, our_filtering, our_selection,
bwasw, blasr))
def get_illumina_comparison_result():
reference_vntrs = load_unique_vntrs_data()
id_to_gene = {
119: 'DRD4',
1220: 'GP1BA',
1221: 'CSTB',
1214: 'MAOA',
1219: 'IL1RN'
}
genes = glob.glob('../Illumina_copy_number/*')
for gene_dir in genes:
print(gene_dir)
files = glob.glob(gene_dir + '/*30x.sam')
gene_name = gene_dir.split('/')[-1]
print(len(files))
mapped_reads = {}
for file_name in files:
copies = file_name.split('_')[-2]
make_bam_and_index(file_name)
base_name = file_name[:-4]
bowtie_bam = bowtie_alignment(base_name + '.fq')
bwa_bam = bwamem_alignment(base_name + '.fq')
original_bam = file_name[:-4] + '.bam'
vntr_id = None
for vid, gname in id_to_gene.items():
if gname == gene_name:
vntr_id = vid
vntr_finder = VNTRFinder(reference_vntrs[vntr_id])
original = count_reads(original_bam)
our_selection = get_our_selected_reads_count(
base_name + '.fq', vntr_finder)
our_filtering = get_our_filtered_reads_count(
base_name + '.fq', vntr_finder)
bwa = count_reads_mapped_to_vntr_in_bamfile(
bwa_bam, reference_vntrs[vntr_id])
bowtie = count_reads_mapped_to_vntr_in_bamfile(
bowtie_bam, reference_vntrs[vntr_id])
mapped_reads[int(copies)] = [
original, our_filtering, our_selection, bwa, bowtie
]
with open(gene_dir + '/result.txt', 'w') as out:
for copies in sorted(mapped_reads.iterkeys()):
original, our_filtering, our_selection, bwa, bowtie = mapped_reads[
copies]
out.write('%s %s %s %s %s %s\n' %
(copies, original, our_filtering, our_selection, bwa,
bowtie))
def create_pacbio_copy_number_variation_references(
pacbio_read_dir='../pacbio_recruitment/set1/'):
from reference_vntr import load_unique_vntrs_data
reference_vntrs = load_unique_vntrs_data()
id_to_gene = {1221: 'CSTB', 1216: 'HIC1', 1215: 'INS'}
repeats = {
'CSTB': range(1, 69),
'HIC1': range(2, 36),
'INS': range(10, 171)
}
for vntr_id in id_to_gene.keys():
for repeat in repeats[id_to_gene[vntr_id]]:
if id_to_gene[vntr_id] == 'INS' and repeat % 5 != 0:
continue
if id_to_gene[vntr_id] == 'CSTB' and repeat % 2 != 0:
continue
outfile = pacbio_read_dir + id_to_gene[vntr_id] + '/' + str(
repeat) + '.fa'
create_reference_region_with_specific_repeats(
reference_vntrs[vntr_id], repeat, outfile, 1000)
def create_pacbio_coverage_data_for_3_genes_and_10_cn(
pacbio_read_dir='../pacbio_coverage_experiment/'):
from reference_vntr import load_unique_vntrs_data
reference_vntrs = load_unique_vntrs_data()
id_to_gene = {1221: 'CSTB', 1216: 'HIC1', 1215: 'INS'}
repeats = {
'CSTB': range(2, 42),
'HIC1': range(2, 22),
'INS': range(10, 110)
}
for vntr_id in id_to_gene.keys():
for repeat in repeats[id_to_gene[vntr_id]]:
if id_to_gene[vntr_id] == 'INS' and repeat % 5 != 0:
continue
if id_to_gene[vntr_id] == 'CSTB' and repeat % 2 != 0:
continue
if id_to_gene[vntr_id] != 'INS':
continue
outfile = pacbio_read_dir + id_to_gene[vntr_id] + '/' + str(
repeat) + '.fa'
create_reference_region_with_specific_repeats(
reference_vntrs[vntr_id], repeat, outfile, 3000)
def create_pacbio_ru_length_data_for_all_vntrs(
pacbio_read_dir='../pacbio_ru_data_for_all_vntrs/'):
from reference_vntr import load_unique_vntrs_data
reference_vntrs = load_unique_vntrs_data()
with open('vntr_complex.txt') as infile:
lines = infile.readlines()
complex_vntrs = [int(r.strip().split()[0]) for r in lines] + [0]
repeat_units = {}
for vntr_id in range(len(reference_vntrs)):
if vntr_id in complex_vntrs:
continue
ru = len(reference_vntrs[vntr_id].pattern)
if ru not in repeat_units.keys():
repeat_units[ru] = []
if len(repeat_units[ru]) >= 4:
continue
repeat_units[ru].append(vntr_id)
import os
for ru in repeat_units.keys():
if len(repeat_units[ru]) < 2:
continue
for vntr_id in repeat_units[ru]:
original_repeats = len(
reference_vntrs[vntr_id].get_repeat_segments())
start = max(3, original_repeats - 10)
for repeat in range(start, start + 21):
if repeat % 5 != 0:
continue
outfile = pacbio_read_dir + str(ru) + '/vntr_id_' + str(
vntr_id) + '_' + str(repeat) + '.fa'
if not os.path.exists(os.path.dirname(outfile)):
os.makedirs(os.path.dirname(outfile))
create_reference_region_with_specific_repeats(
reference_vntrs[vntr_id], repeat, outfile, 1000)
def find_info_by_mapping(sim_dir='simulation_data/', dir_index=0):
reference_vntrs = load_unique_vntrs_data()
id_to_gene = {119: 'DRD4', 1220: 'GP1BA', 1221: 'CSTB', 1214: 'MAOA', 1219: 'IL1RN'}
gene_to_length = {'DRD4': 528, 'GP1BA': 39, 'CSTB': 168, 'MAOA': 30}
clean_up_tmp()
dirs = glob.glob(sim_dir+'/*')
simulation_dir = dirs[dir_index]
files = glob.glob(simulation_dir + '/*')
for fasta_file in files:
if fasta_file.endswith('WGS_30x.fasta'):
gene_name = simulation_dir.split('/')[-1].split('_')[0]
vntr_id = None
for vid, gname in id_to_gene.items():
if gname == gene_name:
vntr_id = vid
ref_vntr = reference_vntrs[vntr_id]
true_reads_file = fasta_file[:-6] + '_true_reads.txt'
simulated_sam_file = fasta_file[:-6] + '.sam'
if not os.path.exists(true_reads_file):
region = [ref_vntr.start_point, ref_vntr.start_point + gene_to_length[gene_name]]
true_reads = get_id_of_reads_mapped_to_vntr_in_samfile(simulated_sam_file, ref_vntr, region=region)
with open(true_reads_file, 'w') as out:
for true_read in true_reads:
out.write('%s\n' % true_read)
else:
with open(true_reads_file) as input:
lines = input.readlines()
true_reads = [line.strip() for line in lines if line.strip() != '']
true_reads_hmm_scores = fasta_file[:-6] + '_t_reads_hmm_score.txt'
false_reads_hmm_scores = fasta_file[:-6] + '_f_reads_hmm_score.txt'
if not os.path.exists(true_reads_hmm_scores):
write_hmm_scores(simulated_sam_file, true_reads_hmm_scores, false_reads_hmm_scores, ref_vntr, true_reads)
for i, parameter in enumerate([10]):
positive_file = fasta_file[:-6] + '_bwa_%s_positive_supplementary_reads.txt' % abs(parameter)
false_negative_file = fasta_file[:-6] + '_bwa_%s_fn_supplementary_reads.txt' % abs(parameter)
if os.path.exists(positive_file) and os.path.exists(false_negative_file):
continue
bwa_alignment_file = '/tmp/_gene%s_' % dir_index + 'bwa_alignment_%s.sam' % i
bwa_alignment(fasta_file, bwa_alignment_file, parameter)
positive_reads, fn_reads = get_positive_and_fn_reads_from_samfile(bwa_alignment_file, ref_vntr, true_reads)
save_reads_stat(positive_file, positive_reads)
save_reads_stat(false_negative_file, fn_reads)
clean_up_tmp()
for i, parameter in enumerate([-0.6, -2]):
if i == 0:
continue
positive_file = fasta_file[:-6] + '_bowtie_%s_positive_supplementary_reads.txt' % abs(parameter)
false_negative_file = fasta_file[:-6] + '_bowtie_%s_fn_supplementary_reads.txt' % abs(parameter)
if os.path.exists(positive_file) and os.path.exists(false_negative_file):
continue
bowtie_alignment_file = '/tmp/_gene%s_' % dir_index + 'bowtie_alignment_%s.sam' % i
bowtie_alignment(fasta_file, bowtie_alignment_file, parameter)
positive_reads, fn_reads = get_positive_and_fn_reads_from_samfile(bowtie_alignment_file, ref_vntr, true_reads)
save_reads_stat(positive_file, positive_reads)
save_reads_stat(false_negative_file, fn_reads)
if gene_name == 'MAOA':
os.system('cp %s /pedigree2/projects/VeNTeR/bowtie_alignment_%s.sam' % (bowtie_alignment_file, i))
clean_up_tmp()
def genotype(args, genotype_parser):
if args.alignment_file is None and args.fasta is None:
print_error(
genotype_parser,
'ERROR: No input specified. Please specify alignment file or fasta file'
)
if args.nanopore:
settings.MAX_ERROR_RATE = 0.3
elif args.pacbio:
settings.MAX_ERROR_RATE = 0.3
else:
settings.MAX_ERROR_RATE = 0.05
if args.threads < 1:
print_error(genotype_parser, 'ERROR: threads cannot be less than 1')
settings.CORES = args.threads
input_file = args.alignment_file if args.alignment_file else args.fasta
input_is_alignment_file = input_file.endswith(
'bam') or input_file.endswith('sam')
working_directory = args.working_directory + '/' if args.working_directory else os.path.dirname(
input_file) + '/'
log_file = working_directory + 'log_%s.log' % os.path.basename(input_file)
log_format = '%(asctime)s %(levelname)s:%(message)s'
logging.basicConfig(format=log_format,
filename=log_file,
level=logging.DEBUG,
filemode='w')
reference_vntrs = load_unique_vntrs_data()
# reference_vntrs = identify_homologous_vntrs(reference_vntrs, 'chr15')
# illumina_targets = [1214, 1220, 1221, 1222, 1223, 1224, 377, 378, 809]
illumina_targets = [532789, 188871, 301645, 600000]
target_vntrs = []
for i in range(len(reference_vntrs)):
if not reference_vntrs[i].is_non_overlapping(
) or reference_vntrs[i].has_homologous_vntr():
continue
target_vntrs.append(i)
if args.vntr_id is not None:
target_vntrs = [int(vid) for vid in args.vntr_id.split(',')]
else:
target_vntrs = illumina_targets
genome_analyzier = GenomeAnalyzer(reference_vntrs, target_vntrs,
working_directory)
if args.pacbio:
if input_is_alignment_file:
genome_analyzier.find_repeat_counts_from_pacbio_alignment_file(
input_file)
else:
genome_analyzier.find_repeat_counts_from_pacbio_reads(
input_file, args.naive)
else:
if args.frameshift:
if valid_vntr_for_frameshift(target_vntrs):
genome_analyzier.find_frameshift_from_alignment_file(
input_file)
else:
genotype_parser.error(
'--frameshift is only available for these IDs: %s' %
settings.FRAMESHIFT_VNTRS)
elif input_is_alignment_file:
genome_analyzier.find_repeat_counts_from_alignment_file(input_file)
else:
genome_analyzier.find_repeat_counts_from_short_reads(input_file)
