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283 lines
12 KiB
Python
283 lines
12 KiB
Python
import os, sys
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import argparse
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from Bio import SeqIO
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import CUB
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from statistics import mean
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from math import ceil, floor
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from tqdm import tqdm
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import matplotlib.pyplot as plt
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import numpy as np
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def get_args():
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parser = argparse.ArgumentParser(
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prog = 'PTL6p1 Script 8: Stat Summary',
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description = "Updated March 31th, 2023 by Auden Cote-L'Heureux"
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)
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parser.add_argument('-i', '--input', type = str, required = True, help = 'Input path to the "Output" folder produced by PhyloToL Part 1. This folder should contain both the "ReadyToGO" and "Intermediate" folders.')
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parser.add_argument('-d', '--databases', type = str, default = '../Databases', help = 'Path to databases folder')
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parser.add_argument('-r', '--r2g_jf', action = 'store_true', help = 'Create ReadyToGo files filtered to only include sequences between the 25th and 75th percentile of silent-site GC content. Please be aware that these are not necessarily the correct or non-contaminant sequences; examine the GC3xENc plots carefully before using these data.')
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#Curate genetic code
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return parser.parse_args()
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def hook_lens(args):
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print('\nGetting average OG lengths in the Hook DB...')
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len_by_og = { }
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for file in os.listdir(args.databases + '/db_OG'):
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if file.endswith('.fasta') and os.path.isfile(args.databases + '/db_OG/' + file.replace('.fasta', '.dmnd')):
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for rec in tqdm(SeqIO.parse(args.databases + '/db_OG/' + file, 'fasta')):
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if rec.id[-10:] not in len_by_og:
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len_by_og.update({ rec.id[-10:] : [] })
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len_by_og[rec.id[-10:]].append(len(str(rec.seq)))
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for og in len_by_og:
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len_by_og[og] = mean(len_by_og[og])
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return len_by_og
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def aa_comp_lengths(args, gcodes):
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print('\nGetting amino acid composition data from ReadyToGo files...')
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r2g_lengths = { }; aa_comp = { }; recid_by_contig_n = { }
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for file in tqdm([f for f in os.listdir(args.input + '/ReadyToGo/ReadyToGo_AA')]):
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if file.endswith('.fasta') and file[:10] in gcodes:
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for rec in SeqIO.parse(args.input + '/ReadyToGo/ReadyToGo_AA/' + file, 'fasta'):
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r2g_lengths.update({ rec.id : len(str(rec.seq)) * 3 })
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fymink = 0; garp = 0; other = 0; total = 0; x = 0
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for char in str(rec.seq):
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if char in 'FYMINKfymink':
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fymink += 1
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elif char in 'GARPgarp':
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garp += 1
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elif char in 'Xx':
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x += 1
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else:
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other += 1
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total += 1
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aa_comp.update({ rec.id : { 'FYMINK' : fymink/total, 'GARP' : garp/total, 'Other' : other/total, 'X' : x} })
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recid_by_contig_n.update({ rec.id.split('Contig_')[-1].split('_')[0] : rec.id })
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print('\nGetting transcript sequence data from original assembled transcript files...')
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transcripts = { }; transcript_id_corr = { }
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for tax in tqdm([f for f in os.listdir(args.input + '/Intermediate/')]):
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if os.path.isdir(args.input + '/Intermediate/' + tax + '/Original'):
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for file in os.listdir(args.input + '/Intermediate/' + tax + '/Original'):
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if file.endswith('_GenBankCDS.fasta'):
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for rec in SeqIO.parse(args.input + '/Intermediate/' + tax + '/Original/' + file, 'fasta'):
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transcripts.update({ rec.id : (file[:10], str(rec.seq)) })
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if rec.id.split('NODE_')[-1].split('_')[0] in recid_by_contig_n:
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transcript_id_corr.update({ recid_by_contig_n[rec.id.split('NODE_')[-1].split('_')[0]] : rec.id})
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return aa_comp, transcripts, r2g_lengths, transcript_id_corr
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def get_nuc_comp(args, gcodes):
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print('\nGetting nucleotide composition data from ReadyToGo files...')
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nuc_comp = { }
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for file in tqdm([f for f in os.listdir(args.input + '/ReadyToGo/ReadyToGo_NTD')]):
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if file.endswith('.fasta') and file[:10] in gcodes:
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cub_out = CUB.CalcRefFasta(args.input + '/ReadyToGo/ReadyToGo_NTD/' + file, gcodes[file[:10]].lower())[0]
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for k in cub_out:
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nuc_comp.update({ k : cub_out[k] })
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return nuc_comp
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def per_seq(args, nuc_comp, aa_comp, all_transcripts, r2g_lengths, transcript_id_corr, og_mean_lens):
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if not os.path.isdir(args.input + '/PerSequenceStatSummaries'):
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os.mkdir(args.input + '/PerSequenceStatSummaries')
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taxa = list(dict.fromkeys([seq[:10] for seq in nuc_comp]))
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for taxon in taxa:
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with open(args.input + '/PerSequenceStatSummaries/' + taxon + '.csv', 'w') as o:
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o.write('Sequence,Taxon,OG,Transcript,TranscriptLength,CDSLength,AvgLengthOGinHook,AmbiguousCodons,GC-Overall,GC1,GC2,GC3,GC3-Degen,ExpWrightENc,ObsWrightENc_6Fold,ObsWrightENc_No6Fold,ObsWeightedENc_6Fold,ObsWeightedENc_No6Fold,FYMINK,GARP,OtherAA,N.Xs\n')
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for rec in nuc_comp:
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if rec[:10] == taxon:
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o.write(rec + ',' + rec[:10] + ',' + rec[-10:])
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try:
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o.write(',' + transcript_id_corr[rec] + ',' + str(len(all_transcripts[transcript_id_corr[rec]][1])))
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except KeyError:
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o.write(',NA,NA')
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o.write(',' + str(r2g_lengths[rec]) + ',' + str(og_mean_lens[rec[-10:]]))
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v = nuc_comp[rec]
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gcs = [str(v.gcOverall), str(v.gc1), str(v.gc2), str(v.gc3), str(v.gc4F)]
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ENc = [str(v.expENc), str(v.obsENc_6F), str(v.obsENc_No6F), str(v.SunENc_6F),str(v.SunENc_No6F)]
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o.write(',' + ','.join([str(v.amb_cdn)] + gcs + ENc))
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o.write(',' + str(aa_comp[rec]['FYMINK']) + ',' + str(aa_comp[rec]['GARP']) + ',' + str(aa_comp[rec]['Other']) + ',' + str(aa_comp[rec]['X']) + '\n')
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def per_tax(args, nuc_comp, aa_comp, all_transcripts, r2g_lengths, gcodes, og_mean_lens):
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taxa = list(dict.fromkeys([seq[:10] for seq in nuc_comp]))
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with open(args.input + '/PerTaxonSummary.csv', 'w') as o:
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o.write('Taxon,TranscriptsInput,Median_GCTranscripts,5-95Perc_GCTranscripts,Median_LenTranscripts,IQR_LenTranscripts,SeqsR2G,OGsR2G,Median_GC3R2G,5Perc_GC3R2G,95Perc_GC3R2G,5-95Perc_GC3R2G,Median_ENcR2G,IQR_ENcR2G,Median_LenR2G,IQR_LenR2G,Prop.G1.5_OGAvg,Prop.L0.5_OGAvg,GeneticCode\n')
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for taxon in taxa:
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try:
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o.write(taxon)
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transcripts = [all_transcripts[seq][1].upper() for seq in all_transcripts if all_transcripts[seq][0] == taxon]
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o.write(',' + str(len(transcripts)))
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transcript_gcs = []
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for transcript in transcripts:
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transcript_gcs.append((transcript.count('G') + transcript.count('C'))/len(transcript))
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transcript_gcs = sorted(transcript_gcs)
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o.write(',' + str(transcript_gcs[floor(len(transcripts)*0.5)]))
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o.write(',' + str(transcript_gcs[floor(len(transcripts)*0.95)] - transcript_gcs[floor(len(transcripts)*0.05)]))
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transcript_lens = sorted([len(transcript) for transcript in transcripts])
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o.write(',' + str(transcript_lens[floor(len(transcripts)*0.5)]))
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o.write(',' + str(transcript_lens[floor(len(transcripts)*0.75)] - transcript_lens[floor(len(transcripts)*0.25)]))
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r2g_ntds = [nuc_comp[seq] for seq in nuc_comp if seq[:10] == taxon]
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o.write(',' + str(len(r2g_ntds)))
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r2g_ogs = list(dict.fromkeys([seq[-10:] for seq in nuc_comp if seq[:10] == taxon]))
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o.write(',' + str(len(r2g_ogs)))
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r2g_gc3s = sorted([seq.gc4F for seq in r2g_ntds])
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o.write(',' + str(r2g_gc3s[floor(len(r2g_ntds)*0.5)]))
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o.write(',' + str(r2g_gc3s[floor(len(r2g_gc3s)*0.05)]))
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o.write(',' + str(r2g_gc3s[floor(len(r2g_gc3s)*0.95)]))
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o.write(',' + str(r2g_gc3s[floor(len(r2g_gc3s)*0.95)] - r2g_gc3s[floor(len(r2g_gc3s)*0.05)]))
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r2g_encs = sorted([seq.obsENc_6F for seq in r2g_ntds])
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o.write(',' + str(r2g_encs[floor(len(r2g_encs)*0.5)]))
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o.write(',' + str(r2g_encs[floor(len(r2g_encs)*0.75)] - r2g_encs[floor(len(r2g_encs)*0.25)]))
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tax_r2g_lens = sorted([r2g_lengths[seq] for seq in r2g_lengths if seq[:10] == taxon])
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o.write(',' + str(tax_r2g_lens[floor(len(tax_r2g_lens)*0.5)]))
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o.write(',' + str(tax_r2g_lens[floor(len(tax_r2g_lens)*0.75)] - tax_r2g_lens[floor(len(tax_r2g_lens)*0.25)]))
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prop_len_g = len([seq for seq in r2g_lengths if seq[:10] == taxon and r2g_lengths[seq] > 4.5 * og_mean_lens[seq[-10:]]])/len(tax_r2g_lens)
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prop_len_l = len([seq for seq in r2g_lengths if seq[:10] == taxon and r2g_lengths[seq] < 1.5 * og_mean_lens[seq[-10:]]])/len(tax_r2g_lens)
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o.write(',' + str(prop_len_g) + ',' + str(prop_len_l))
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o.write(',' + gcodes[taxon] + '\n')
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except:
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pass
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def r2g_jf(args, nuc_comp, gcodes):
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#Q: should there be an maximum IQR cutoff at which we do NOT produce a file here?
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if not os.path.isdir(args.input + '/ReadyToGo/ReadyToGo_NTD_JF'):
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os.mkdir(args.input + '/ReadyToGo/ReadyToGo_NTD_JF')
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if not os.path.isdir(args.input + '/ReadyToGo/ReadyToGo_AA_JF'):
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os.mkdir(args.input + '/ReadyToGo/ReadyToGo_AA_JF')
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for file in os.listdir(args.input + '/ReadyToGo/ReadyToGo_NTD'):
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if file.endswith('.fasta') and file[:10] in gcodes:
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taxon = file[:10]
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r2g_ntds = [nuc_comp[seq] for seq in nuc_comp if seq[:10] == taxon]
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r2g_gc3s = sorted([seq.gc4F for seq in r2g_ntds])
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with open(args.input + '/ReadyToGo/ReadyToGo_NTD_JF/' + file.replace('.fasta', '.JF.fasta'), 'w') as o:
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for rec in SeqIO.parse(args.input + '/ReadyToGo/ReadyToGo_NTD/' + file, 'fasta'):
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if nuc_comp[rec.id].gc4F > r2g_gc3s[floor(len(r2g_gc3s)*0.25)] and nuc_comp[rec.id].gc4F < r2g_gc3s[floor(len(r2g_gc3s)*0.75)]:
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o.write('>' + rec.id + '\n' + str(rec.seq) + '\n\n')
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with open(args.input + '/ReadyToGo/ReadyToGo_AA_JF/' + file.replace('.fasta', '.JF.fasta').replace('NTD', 'AA'), 'w') as o:
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for rec in SeqIO.parse(args.input + '/ReadyToGo/ReadyToGo_AA/' + file.replace('NTD', 'AA'), 'fasta'):
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if nuc_comp[rec.id].gc4F > r2g_gc3s[floor(len(r2g_gc3s)*0.25)] and nuc_comp[rec.id].gc4F < r2g_gc3s[floor(len(r2g_gc3s)*0.75)]:
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o.write('>' + rec.id + '\n' + str(rec.seq) + '\n\n')
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def plot_jf(args, nuc_comp):
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if not os.path.isdir(args.input + '/GC3xENc_Plots'):
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os.mkdir(args.input + '/GC3xENc_Plots')
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taxa = list(dict.fromkeys([rec[:10] for rec in nuc_comp]))
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gc3_null = [0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100]
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enc_null = [31, 31.5958, 32.2032, 32.8221, 33.4525, 34.0942, 34.7471, 35.411, 36.0856, 36.7707, 37.4659, 38.1707, 38.8847, 39.6074, 40.3381, 41.0762, 41.8208, 42.5712, 43.3264, 44.0854, 44.8471, 45.6102, 46.3735, 47.1355, 47.8949, 48.65, 49.3991, 50.1406, 50.8725, 51.593, 52.3, 52.9916, 53.6656, 54.32, 54.9525, 55.561, 56.1434, 56.6975, 57.2211, 57.7124, 58.1692, 58.5898, 58.9723, 59.3151, 59.6167, 59.8757, 60.0912, 60.2619, 60.3873, 60.4668, 60.5, 60.4668, 60.3873, 60.2619, 60.0912, 59.8757, 59.6167, 59.3151, 58.9723, 58.5898, 58.1692, 57.7124, 57.2211, 56.6975, 56.1434, 55.561, 54.9525, 54.32, 53.6656, 52.9916, 52.3, 51.593, 50.8725, 50.1406, 49.3991, 48.65, 47.8949, 47.1355, 46.3735, 45.6102, 44.8471, 44.0854, 43.3264, 42.5712, 41.8208, 41.0762, 40.3381, 39.6074, 38.8847, 38.1707, 37.4659, 36.7707, 36.0856, 35.411, 34.7471, 34.0942, 33.4525, 32.8221, 32.2032, 31.5958, 31]
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for taxon in taxa:
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comp_data = [(nuc_comp[rec].gc4F, nuc_comp[rec].obsENc_6F) for rec in nuc_comp if rec[:10] == taxon]
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plt.figure()
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plt.plot(np.array(gc3_null), np.array(enc_null), color = 'black', linewidth=2)
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plt.scatter(np.array([val[0] for val in comp_data]), np.array([val[1] for val in comp_data]), s = 1)
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plt.xlabel("GC content (3rd pos, 4-fold sites)")
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plt.ylabel("Observed Wright ENc (6 Fold)")
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plt.savefig(args.input + '/GC3xENc_Plots/' + taxon + '.png')
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if __name__ == "__main__":
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args = get_args()
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valid_codes = ['universal', 'blepharisma', 'chilodonella', 'condylostoma', 'euplotes', 'peritrich', 'vorticella', 'mesodinium', 'tag', 'tga', 'taa', 'none']
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gcodes = { }
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if os.path.isfile(args.input + '/Intermediate/gcode_output.tsv'):
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for line in open(args.input + '/Intermediate/gcode_output.tsv'):
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if len(line.split('\t')) == 5 and line.split('\t')[4].strip().lower() in valid_codes:
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gcodes.update({ line.split('\t')[0] : line.split('\t')[4].strip() })
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elif line.split('\t')[4].strip().lower() != '':
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print('\nInvalid genetic code assignment for taxon ' + line.split('\t')[0] + '. Skipping this taxon in script 6 (summary statistics)\n')
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else:
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print('\nGenetic code assignment file (Output/Intermediate/gcode_output.tsv) not found. Quitting script 6 (summary statistics).\n')
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exit()
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aa_comp, transcripts, r2g_lengths, transcript_id_corr = aa_comp_lengths(args, gcodes)
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nuc_comp = get_nuc_comp(args, gcodes)
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og_mean_lens = hook_lens(args)
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per_tax(args, nuc_comp, aa_comp, transcripts, r2g_lengths, gcodes, og_mean_lens)
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per_seq(args, nuc_comp, aa_comp, transcripts, r2g_lengths, transcript_id_corr, og_mean_lens)
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if args.r2g_jf:
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r2g_jf(args, nuc_comp, gcodes)
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plot_jf(args, nuc_comp)
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