''' Written March 2023 by Elinor (esterner27@gmail.com) to plot length, coverage and GC of assembled transcripts This script will rename the spades output to new names in the txt file, then iterate through them all and gather GC, length and coverage. With that data, it plots R scripts Input: Directory of directories output by rnaSpades OR folder called Renamed_assembled_files of previously renamed files (if this is the case, put -r or --renamed in the command line) txt file of LKH number and new names formatted like this: LKHxxx\tLKHxxx-10_digit_code-descriptor_of_taxon To run if your files are already renamed: python assess_transcriptomes.py python assess_transcriptomes.py --renamed 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:\ntxt file of tab separated LKH number, ten digit code and taxon info (taxonomy, lifestage, etc).\nAND\nfolder of the folders output by spades named with LKH number \n(LKH999 or WTALKH999)\nOR\ndirectory of renamed assemblies 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 \n\nOptional parameters:\n\n-r or --renamed if your assemblies are already renamed to this format: ten_digit_code_assembledTranscript.fasta\n\n-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. renamed = False if('--help' in sys.argv or '-h' in sys.argv):#check for help function in command line script_help() exit() if ('--renamed'in sys.argv or '-r' in sys.argv):#check for renamed parameter renamed = True try: if('--renamed' in sys.argv): input_dir = sys.argv[sys.argv.index('--renamed') + 1] else: input_dir = sys.argv[sys.argv.index('-r') + 1] except IndexError: print('\nSomething went wrong went parsing the arguments. Did you input a directory of assemblies?\n') if ('--raw' in sys.argv or '-w' in sys.argv):#check for renamed parameter renamed = False try: if('--raw' in sys.argv): input_dir = sys.argv[sys.argv.index('--raw') + 1] else: input_dir = sys.argv[sys.argv.index('-r') + 1] except IndexError: print('\nSomething went wrong went parsing the arguments. Did you input a directory of assemblies?\n') make_dirs(renamed, input_dir) def make_dirs(renamed, input_dir): Path(f'plots').mkdir(parents=True, exist_ok=True)#makes output folder for r plots if renamed == True: assess_transcriptomes(input_dir)#skip renaming funtion if theyre already renamed elif renamed == False: Path(f'Renamed_assembled_files').mkdir(parents=True, exist_ok=True)#makes output folder for renamed fasta files #print('renaming') #print(input_dir) rename_assembled_transcriptomes(input_dir)#send to renaming function if they're not renamed yet. else: print('\nplease specify if your files are already renamed (--renamed) or if they are not yet renamed (--raw)\n') def rename_assembled_transcriptomes(input_dir):#this function grabs and renames assembled transcripts files to the format required for this script and for phylotolv6 part1 names = { line.split('\t')[0].strip() : line.split('\t')[1].strip() for line in open('new_names.txt')}#make dictionary of original name: new name to replace for dir in os.listdir(input_dir): for old_name, new_name in names.items(): if(old_name in dir): if(os.path.isfile(f'{input_dir}/{dir}/transcripts.fasta')):#go into spades output directory print(f'{old_name} is being renamed to {new_name}\n') os.system(f'cp {input_dir}/{dir}/transcripts.fasta Renamed_assembled_files/{new_name}_assembledTranscripts.fasta')#rename and copy to renamed_assembled files input_dir = 'Renamed_assembled_files' assess_transcriptomes(input_dir) 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 lkh = taxon_info[0] ten_digit_code = taxon_info[1] taxon = taxon_info[2] # 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}, {lkh}, {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.txt file to get info on the taxon with open('new_names.txt', 'r') as o: cell_info = o.readlines() for line in cell_info: if '_'.join(file.split('_')[0:3]) in line: lkh = line.split('\t')[0] ten_digit_code = line.split('\t')[1] taxon = line.split('\t')[2] try: return lkh, ten_digit_code, taxon except UnboundLocalError: print(f'no taxon information in new_names.txt for {file}') print('done with getting taxon info') 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, lkh, 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}')#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 = {} get_args()