# HG changeset patch # User kaymccoy # Date 1481493685 18000 # Node ID 5ff57a3d0af2b71de6b9df95182fb4649d7b0687 # Parent 359b7cc9ad9466bcaa3b98ea02389074bca2ddd7 Uploaded diff -r 359b7cc9ad94 -r 5ff57a3d0af2 aggregate.py --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/aggregate.py Sun Dec 11 17:01:25 2016 -0500 @@ -0,0 +1,297 @@ +# A translation of aggregate.pl into python! For analysis of Tn-Seq. +# This script requires BioPython just like calc_fitness.py, so you need it installed along with its dependencies if you want to run these scripts on your own. +# How to install BioPython and a list of its dependencies can be found here: http://biopython.org/DIST/docs/install/Installation.html +# K. McCoy + + + + + + + + + +##### ARGUMENTS ##### + +def print_usage(): + print "Aggregate.py's usage is as follows:" + "\n\n" + print "\033[1m" + "Required" + "\033[0m" + "\n" + print "-o" + "\t\t" + "Output file for aggregated data." + "\n" + print "\n" + print "\033[1m" + "Optional" + "\033[0m" + "\n" + print "-c" + "\t\t" + "Check for missing genes in the data set - provide a reference genome in genbank format. Missing genes will be sent to stdout." + "\n" + print "-m" + "\t\t" + "Place a mark in an extra column for this set of genes. Provide a file with a list of genes seperated by newlines." + "\n" + print "-x" + "\t\t" + "Cutoff: Don't include fitness scores with average counts (c1+c2)/2 < x (default: 0)" + "\n" + print "-b" + "\t\t" + "Bottleneck value: The percentage of insertions randomly lost, which will be discounted for all genes (for example, 20% would be entered as 0.20; default 0.0)" + "\n" + print "-f" + "\t\t" + "An in-between file carrying information on the blank count found from calc_fitness or consol_fitness; one of two ways to pass a blank count to this script" + "\n" + print "-w" + "\t\t" + "Use weighted algorithm to calculate averages, variance, sd, se" + "\n" + print "-l" + "\t\t" + "Weight ceiling: maximum value to use as a weight (default: 999,999)" + "\n" + print "\n" + print "All remainder arguements will be treated as fitness files (those files created by calc_fitness.py)" + "\n" + print "\n" + +import argparse +parser = argparse.ArgumentParser() +parser.add_argument("-o", action="store", dest="summary") +parser.add_argument("-c", action="store", dest="find_missing") +parser.add_argument("-m", action="store", dest="marked") +parser.add_argument("-x", action="store", dest="cutoff") +parser.add_argument("-b", action="store", dest="blank_pc") +parser.add_argument("-f", action="store", dest="blank_file") +parser.add_argument("-w", action="store", dest="weighted") +parser.add_argument("-l", action="store", dest="weight_ceiling") +parser.add_argument("fitnessfiles", nargs=argparse.REMAINDER) + +arguments = parser.parse_args() + +if not arguments.summary: + print "\n" + "You are missing a value for the -o flag. " + print_usage() + quit() + +if not arguments.fitnessfiles: + print "\n" + "You are missing fitness file(s); these should be entered immediately after all the flags. " + print_usage() + quit() + +# 999,999 is a trivial placeholder number + +if (not arguments.weight_ceiling): + arguments.weight_ceiling = 999999 + +# Cutoff exists to discard positions with a low number of counted transcripts, because their fitness may not be as accurate - for the same reasoning that studies with low sample sizes can be innacurate. + +if (not arguments.cutoff): + arguments.cutoff = 0 + +# Gets information from the txt output file of calc_fit / consol, if inputted + +if arguments.blank_file: + with open(arguments.blank_file) as file: + blank_pc = file.read().splitlines() + arguments.blank_pc = float(blank_pc[0].split()[1]) + +if (not arguments.blank_pc): + arguments.blank_pc = 0 + + + + + +##### SUBROUTINES ##### + +# A subroutine that calculates the average, variance, standard deviation (sd), and standard error (se) of a group of scores; for use when aggregating scores by gene later on + +import math +def unweighted_average(scores): + sum = 0 + num = 0 + i = 0 + while i < len(scores): + if not scores[i]: + scores[i] = 0.0 + sum += float(scores[i]) + num += 1 + i += 1 + average = sum/num + xminusxbars = 0 + while i < len(scores): + xminusxbars += (float(scores[i]) - average)**2 + if num <= 1: + variance = 0 + else: + variance = xminusxbars/(num-1) + sd = math.sqrt(variance) + se = sd / math.sqrt(num) + return (average, variance, sd, se) + +# A subroutine that calculates the weighted average, variance, standard deviation (sd), and standard error (se) of a group of scores; the weights come from the number of reads each insertion location has +# For use when aggregating scores by gene later on, if the weighted argument is called + +def weighted_average(scores,weights): + sum = 0 + weighted_average = 0 + weighted_variance = 0 + top = 0 + bottom = 0 + i = 0 + while i < len(weights): + if not scores[i]: + scores[i] = 0.0 + top += float(weights[i])*float(scores[i]) + bottom += float(weights[i]) + i += 1 + if bottom == 0: + return 0 + weighted_average = top/bottom + top = 0 + bottom = 0 + i = 0 + while i < len(weights): + top += float(weights[i]) * (float(scores[i]) - weighted_average)**2 + bottom += float(weights[i]) + i += 1 + weighted_variance = top/bottom + weighted_stdev = math.sqrt(weighted_variance) + weighted_stder = weighted_stdev/math.sqrt(len(scores)) + return (weighted_average, weighted_variance, weighted_stdev, weighted_stder) + + + + + + + + + + +##### AGGREGATION / CALCULATIONS ##### + +#Reads the genes which should be marked in the final aggregate file into an array + +import os.path +if arguments.marked: + with open(arguments.marked) as file: + marked_set = file.read().splitlines() + +#Creates a dictionary of dictionaries to contain a summary of all genes and their fitness values +#The fitness values and weights match up, so that the weight of gene_summary[locus]["w"][2] would be gene_summary[locus]["s"][2] + +import csv +gene_summary = {} +for eachfile in arguments.fitnessfiles: + with open(eachfile) as csvfile: + lines = csv.reader(csvfile) + for line in lines: + locus = line[9] + w = line[12] + if w == 'nW': + continue + if not w: + w == 0 + c1 = float(line[2]) + c2 = float(line[3]) + avg = (c1+c2)/2 + if avg < float(arguments.cutoff): + continue + if avg > float(arguments.weight_ceiling): + avg = arguments.weight_ceiling + if locus not in gene_summary: + gene_summary[locus] = {"w" : [], "s": []} + gene_summary[locus]["w"].append(w) + gene_summary[locus]["s"].append(avg) + +#If finding any missing gene loci is requested in the arguments, starts out by loading all the known features from a genbank file + +from Bio import SeqIO +if (arguments.find_missing): + output = [["locus","mean","var","sd","se","gene","Total","Blank","Not Blank","Blank Removed","M\n"]] + handle = open(arguments.find_missing, "rU") + for record in SeqIO.parse(handle, "genbank"): + refname = record.id + features = record.features + handle.close() + +#Goes through the features to find which are genes + + for feature in features: + gene = "" + if feature.type == "gene": + locus = "".join(feature.qualifiers["locus_tag"]) + if "gene" in feature.qualifiers: + gene = "".join(feature.qualifiers["gene"]) + else: + continue + +#Goes through the fitness scores of insertions within each gene, and removes whatever % of blank fitness scores were requested along with their corresponding weights + + sum = 0 + num = 0 + avgsum = 0 + blank_ws = 0 + i = 0 + if locus in gene_summary.keys(): + for w in gene_summary[locus]["w"]: + if float(w) == 0: + blank_ws += 1 + else: + sum += float(w) + num += 1 + count = num + blank_ws + removed = 0 + to_remove = int(float(arguments.blank_pc)*count) + if blank_ws > 0: + i = 0 + while i < len(gene_summary[locus]["w"]): + w = gene_summary[locus]["w"][i] + if removed == to_remove: + break + if float(w) == 0: + del gene_summary[locus]["w"][i] + del gene_summary[locus]["s"][i] + removed += 1 + i -= 1 + i += 1 + +#If all the fitness values within a gene are empty, sets mean/var to 0.10 and Xs out sd/se; marks the gene if that's requested + + if num == 0: + if (arguments.marked and locus in marked_set): + output.append([locus, "0.10", "0.10", "X", "X", gene, count, blank_ws, num, removed, "M", "\n"]) + else: + output.append([locus, "0.10", "0.10", "X", "X", gene, count, blank_ws, num, removed, "\n"]) + +#Otherwise calls average() or weighted_average() to find the aggregate w / count / standard deviation / standard error of the insertions within each gene; marks the gene if that's requested + + else: + if not arguments.weighted: + (average, variance, stdev, stderr) = unweighted_average(gene_summary[locus]["w"]) + else: + (average, variance, stdev, stderr) = weighted_average(gene_summary[locus]["w"],gene_summary[locus]["s"]) + if (arguments.marked and locus in marked_set): + output.append([locus, average, variance, stdev, stderr, gene, count, blank_ws, num, removed, "M", "\n"]) + else: + output.append([locus, average, variance, stdev, stderr, gene, count, blank_ws, num, removed, "\n"]) + +#If a gene doesn't have any insertions, sets mean/var to 0.10 and Xs out sd/se, plus leaves count through removed blank because there were no reads. + + else: + if (arguments.marked and locus in marked_set): + output.append([locus, "0.10", "0.10", "X", "X", gene, "", "", "", "", "M", "\n"]) + else: + output.append([locus, "0.10", "0.10", "X", "X", gene, "", "", "", "", "\n"]) + +#Writes the aggregated fitness file + + with open(arguments.summary, "wb") as csvfile: + writer = csv.writer(csvfile) + writer.writerows(output) + +#If finding missing genes is not requested, just finds the aggregate w / count / standard deviation / standard error of the insertions within each gene, and writes them to a file, plus marks the genes requested +#This is never called through Galaxy since finding missing genes is just better than not finding them. + +else: + output = [["Locus","W","Count","SD","SE","M\n"]] + for gene in gene_summary.keys(): + sum = 0 + num = 0 + average = 0 + if "w" not in gene_summary[gene]: + continue + for i in gene_summary[gene]["w"]: + sum += i + num += 1 + average = sum/num + xminusxbars = 0 + for i in w: + xminusxbars += (i-average)**2 + if num > 1: + sd = math.sqrt(xminusxbars/(num-1)) + se = sd / math.sqrt(num) + if (arguments.marked and locus in marked_set): + output.append([gene, average, num, sd, se, "M", "\n"]) + else: + output.append([gene, average, num, sd, se, "\n"]) + with open(arguments.summary, "wb") as csvfile: + writer = csv.writer(csvfile) + writer.writerows(output) \ No newline at end of file