''' Written March 2023 by Elinor (esterner27@gmail.com) to plot length, coverage and GC of assembled transcripts This script will iterate through all assembled files (named as 10 digit code _assembledTranscripts) with and gather GC, length and coverage. With that data, it plots R scripts Input: Folder called Renamed_assembled_files of previously renamed files (if this is the case, put -r or --renamed in the command line) tsv file of LKH number and new names formatted like this: 10_digit_code\tdescriptor_of_taxon called new_names.tsv To run: python assess_transcriptomes.py -input Output: csv file of length, GC, coverage of each transcript, and multiple R plots, faceted by taxon and a csv file of data. It plots GC by length, and distributions of coverage, length and GC ''' import os import sys from pathlib import Path import sys from Bio.SeqUtils import GC from Bio import SeqIO def script_help(): print('\nThis script grabs and plots GC, length and coverage of transcriptomes. \n\nInput:\ntsv file of tab separated ten digit code and taxon info (taxonomy, lifestage, etc).\nAND\n\ndirectory of assemblies renamed in this format: ten_digit_code_assembledTranscripts.fasta\n\nOutput is multiple R plots faceted by taxon and a csv file of data. \n\nIt plots GC by length, and distributions of coverage, length and GC.\n\n To run: \n\n\tpython assess_transcriptomes.py -i \n\nRun -h or --help for this message\n\n') def get_args(): #this parses user arguments. Checks if the files are renamed already or not (--renamed or --raw), and gets the directory of those files. if('--help' in sys.argv or '-h' in sys.argv):#check for help function in command line script_help() exit() if ('--input'in sys.argv or '-i' in sys.argv):#check for renamed parameter renamed = True try: if('--input' in sys.argv): input_dir = sys.argv[sys.argv.index('--input') + 1] else: input_dir = sys.argv[sys.argv.index('-i') + 1] except IndexError: print('\nSomething went wrong went parsing the arguments. Did you input a directory of assemblies?\n') make_dirs(input_dir) def make_dirs(input_dir): Path(f'plots').mkdir(parents=True, exist_ok=True)#makes output folder for r plots assess_transcriptomes(input_dir)#skip renaming funtion if theyre already renamed def assess_transcriptomes(input_dir): #iterate through renamed fasta files for file in os.listdir(input_dir): if file.endswith('fasta'): print(f'gathering data from {file}..\n') data = []#initiate list of data per LKH records = list(SeqIO.parse(f'{input_dir}/{file}', 'fasta'))#parse the fasta files taxon_info = get_taxon_info(file)#send to get_taxon_info function #parse output from that function ten_digit_code = taxon_info[0] taxon = taxon_info[1] # extract all data for each transcript for record in records: gc = GC(record.seq) length = len(record.seq) iden = record.id cov = record.id.split('_')[5] transcript_data = f'{iden}, {length}, {gc}, {cov}, {ten_digit_code}, {taxon}'#list of data per transcriptome data.append(transcript_data)#data for each LKH all_data.update({file.split('_assembledTranscripts')[0]: data})#make dict of ten digit code: all information write_to_file() def get_taxon_info(file):#parse the info in the new_names.tsv file to get info on the taxon with open('new_names.tsv', 'r') as o: cell_info = o.readlines() for line in cell_info: if '_'.join(file.split('_')[0:3]) in line: ten_digit_code = line.split('\t')[0] taxon = line.split('\t')[1] try: return ten_digit_code, taxon except UnboundLocalError: print(f'no taxon information in new_names.tsv for {file}') print('done getting taxon info from new_names.tsv') def write_to_file(): with open('assembly_assessment.csv', 'w') as o:#create output csv print('writing data to assembly_assessment.csv\n') o.write('seqID, length, GC, cov, ten_digit_code, taxon_info\n')#write header for lkh, data in all_data.items(): for transcript in data: o.write(f'{transcript.strip()}\n')#write each line plot_assessment() def plot_assessment(): print('Plotting your data\n') os.system('Rscript plot_assemblies.R') for file in os.listdir(): if file.endswith('.png') or file.endswith('.pdf'): os.system(f'mv {file} plots/{file}') print('\n\n\n\nDone! All your plots are saved to the folder called plots\n\n') if __name__ == '__main__': all_data = {} try: f = open("new_names.tsv") except FileNotFoundError: print('\n\n Did you include a tsv file of your cells? It should be called new_names.tsv and formatted like this: 10_digit_code\tdescriptor_of_taxon\n\n') exit() else: get_args()