comparison aggregate.py @ 4:9dd574c52765 draft

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author kaymccoy
date Thu, 11 Aug 2016 18:30:23 -0400
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3:07ef2716dc03 4:9dd574c52765
1 # A translation of aggregate.pl into python! For analysis of Tn-Seq.
2 # 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.
3 # How to install BioPython and a list of its dependencies can be found here: http://biopython.org/DIST/docs/install/Installation.html
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14 ##### ARGUMENTS #####
15
16 # Prints basic instructions on / options for using this code; called when the user forgets to enter an output file or fitness file(s)
17
18 def print_usage():
19 print "Aggregate.py's usage is as follows:" + "\n\n"
20 print "\033[1m" + "Required" + "\033[0m" + "\n"
21 print "-o" + "\t\t" + "Output file for aggregated data." + "\n"
22 print "\n"
23 print "\033[1m" + "Optional" + "\033[0m" + "\n"
24 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"
25 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"
26 print "-x" + "\t\t" + "Cutoff: Don't include fitness scores with average counts (c1+c2)/2 < x (default: 0)" + "\n"
27 print "-b" + "\t\t" + "Blanks: Exclude -b % of blank fitness scores (scores where c2 = 0) (default: 0 = 0%)" + "\n"
28 print "-w" + "\t\t" + "Use weighted algorithm to calculate averages, variance, sd, se" + "\n"
29 print "-l" + "\t\t" + "Weight ceiling: maximum value to use as a weight (default: 999,999)" + "\n"
30 print "\n"
31 print "All remainder arguements will be treated as fitness files (those files created by calc_fitness.py)" + "\n"
32 print "\n"
33
34 # Turns the arguments entered in the command line into variables so that they can actually be called.
35
36 import argparse
37 parser = argparse.ArgumentParser()
38 parser.add_argument("-o", action="store", dest="summary")
39 parser.add_argument("-c", action="store", dest="find_missing")
40 parser.add_argument("-m", action="store", dest="marked")
41 parser.add_argument("-x", action="store", dest="cutoff")
42 parser.add_argument("-b", action="store", dest="blank_pc")
43 parser.add_argument("-w", action="store", dest="weighted")
44 parser.add_argument("-l", action="store", dest="weight_ceiling")
45 parser.add_argument("fitnessfiles", nargs=argparse.REMAINDER)
46
47 #Shortens the names of those variables created from the arguments - from parser.parse_args().ref_genome to arguments.ref_genome, for example.
48
49 arguments = parser.parse_args()
50
51 #Checks that the required arguments have been entered; if not informs the user they need to provide a name for the output file / the fitness file(s) and print's aggregate.py's usage
52
53 if not arguments.summary:
54 print "\n" + "You are missing a value for the -o flag. "
55 print_usage()
56 quit()
57
58 if not arguments.fitnessfiles:
59 print "\n" + "You are missing fitness file(s); these should be entered immediately after all the flags. "
60 print_usage()
61 quit()
62
63 #Sets the maximum weight of a fitness value, if that wasn't specified in the command line.
64
65 if (not arguments.weight_ceiling):
66 arguments.max_weight = 999999
67
68 #Sets the cutoff to a default value of 0 if it wasn't specified in the command line.
69 #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.
70
71 if (not arguments.cutoff):
72 arguments.cutoff = 0
73
74 #Sets the % of blank fitness values to exclude to a default value of 0 if it wasn't specified in the command line. This can be found in the 2nd output of calc_fitness.
75
76 if (not arguments.blank_pc):
77 arguments.blank_pc = 0
78
79 # gets blank_pc from the output file of calc_fit / consol
80
81 if arguments.blank_pc:
82 with open(arguments.blank_pc) as file:
83 blank_pc = file.read().splitlines()
84 arguments.blank_pc = float(blank_pc[1].split()[1])
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91
92 ##### SUBROUTINES #####
93
94 #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
95
96 import math
97 def average(scores):
98 sum = 0
99 num = 0
100
101 #Finds the average of the scores
102
103 for i in scores:
104 sum += i
105 num += 1
106 average = sum/num
107
108 #Finds the variance of the scores
109
110 xminusxbars = 0
111 for i in scores:
112 xminusxbars += (i - average)**2
113 variance = xminusxbars/(num-1)
114
115 #Finds the standard deviation and standard error of the scores; then the average / variance / standard deviation / standard error are returned
116
117 sd = math.sqrt(variance)
118 se = sd / math.sqrt(num)
119 return (average, variance, sd, se)
120
121 #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
122 #For use when aggregating scores by gene later on, if the weighted argument is called
123
124 def weighted_average(scores,weights):
125 sum = 0
126 weighted_average = 0
127 weighted_variance = 0
128 top = 0
129 bottom = 0
130 i = 0
131
132 #Finds weighted average of the scores
133
134 while i < len(weights):
135 if not scores[i]:
136 scores[i] = 0.0
137 top += float(weights[i])*float(scores[i])
138 bottom += float(weights[i])
139 i += 1
140 if bottom == 0:
141 return 0
142 weighted_average = top/bottom
143
144 #Finds weighted variance of the scores
145
146 top = 0
147 bottom = 0
148 i = 0
149 while i < len(weights):
150 top += float(weights[i]) * (float(scores[i]) - weighted_average)**2
151 bottom += float(weights[i])
152 i += 1
153 weighted_variance = top/bottom
154
155 #Finds weighted standard deviation and standard error of the scores; then the weighted average / variance / standard deviation / standard error are returned
156
157 weighted_stdev = math.sqrt(weighted_variance)
158 weighted_stder = weighted_stdev/math.sqrt(len(scores))
159 return (weighted_average, weighted_variance, weighted_stdev, weighted_stder)
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169
170 ##### AGGREGATION / CALCULATIONS #####
171
172 #Reads the genes which should be marked in the final aggregate file into an array
173
174 import os.path
175 if arguments.marked:
176 with open(arguments.marked) as file:
177 marked_set = file.read().splitlines()
178
179 #Creates a dictionary of dictionaries to contain a summary of all genes and their fitness values
180 #Each gene is its own dictionary with w and s as keys for the various fitnesses and weights of the insertion locations within those genes respectively
181 #The fitness values and weights match up, so that the weight of gene_summary[locus]["w"][2] would be gene_summary[locus]["s"][2]
182
183 import csv
184 gene_summary = {}
185 for eachfile in arguments.fitnessfiles:
186 with open(eachfile) as csvfile:
187 lines = csv.reader(csvfile)
188 for line in lines:
189 locus = line[9]
190 w = line[12]
191 if w == 'nW':
192 continue
193 if not w:
194 w == 0
195 c1 = float(line[2])
196 c2 = float(line[3])
197 avg = (c1+c2)/2
198 if avg < float(arguments.cutoff):
199 continue
200 if avg > float(arguments.weight_ceiling):
201 avg = arguments.weight_ceiling
202 if locus not in gene_summary:
203 gene_summary[locus] = {"w" : [], "s": []}
204 gene_summary[locus]["w"].append(w)
205 gene_summary[locus]["s"].append(avg)
206
207 #If finding any missing gene loci is requested in the arguments, starts out by loading all the known features from a genbank file
208
209 from Bio import SeqIO
210 if (arguments.find_missing):
211 output = [["locus","mean","var","sd","se","gene","Total","Blank","Not Blank","Blank Removed","M\n"]]
212 handle = open(arguments.find_missing, "rU")
213 for record in SeqIO.parse(handle, "genbank"):
214 refname = record.id
215 features = record.features
216 handle.close()
217
218 #Goes through the features to find which are genes
219
220 for feature in features:
221 gene = ""
222 if feature.type == "gene":
223 locus = "".join(feature.qualifiers["locus_tag"])
224 if "gene" in feature.qualifiers:
225 gene = "".join(feature.qualifiers["gene"])
226 else:
227 continue
228
229 #Goes through the fitness scores of insertions within each gene, and removes whatever % of blank fitness scores were requested along with their corresponding weights
230
231 sum = 0
232 num = 0
233 avgsum = 0
234 blank_ws = 0
235 i = 0
236 if locus in gene_summary.keys():
237 for w in gene_summary[locus]["w"]:
238 if float(w) == 0:
239 blank_ws += 1
240 else:
241 sum += float(w)
242 num += 1
243 count = num + blank_ws
244 removed = 0
245 to_remove = int(float(arguments.blank_pc)*count)
246 if blank_ws > 0:
247 i = 0
248 while i < len(gene_summary[locus]["w"]):
249 if removed == to_remove:
250 break
251 if not w:
252 del gene_summary[locus]["w"][i]
253 del gene_summary[locus]["s"][i]
254 removed += 1
255 i -= 1
256 i += 1
257
258 #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
259
260 if num == 0:
261 if (arguments.marked and locus in marked_set):
262 output.append([locus, "0.10", "0.10", "X", "X", gene, count, blank_ws, num, removed, "M", "\n"])
263 else:
264 output.append([locus, "0.10", "0.10", "X", "X", gene, count, blank_ws, num, removed, "\n"])
265
266 #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
267
268 else:
269 if not arguments.weighted:
270 (average, variance, stdev, stderr) = average(gene_summary[locus]["w"])
271 else:
272 (average, variance, stdev, stderr) = weighted_average(gene_summary[locus]["w"],gene_summary[locus]["s"])
273 if (arguments.marked and locus in marked_set):
274 output.append([locus, average, variance, stdev, stderr, gene, count, blank_ws, num, removed, "M", "\n"])
275 else:
276 output.append([locus, average, variance, stdev, stderr, gene, count, blank_ws, num, removed, "\n"])
277
278 #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.
279
280 else:
281 if (arguments.marked and locus in marked_set):
282 output.append([locus, "0.10", "0.10", "X", "X", gene, "", "", "", "", "M", "\n"])
283 else:
284 output.append([locus, "0.10", "0.10", "X", "X", gene, "", "", "", "", "\n"])
285
286 #Writes the aggregated fitness file
287
288 with open(arguments.summary, "wb") as csvfile:
289 writer = csv.writer(csvfile)
290 writer.writerows(output)
291
292 #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
293 #This is never called through Galaxy since finding missing genes is better than not finding them.
294
295 else:
296 output = [["Locus","W","Count","SD","SE","M\n"]]
297 for gene in gene_summary.keys():
298 sum = 0
299 num = 0
300 average = 0
301 if "w" not in gene_summary[gene]:
302 continue
303 for i in gene_summary[gene]["w"]:
304 sum += i
305 num += 1
306 average = sum/num
307 xminusxbars = 0
308 for i in w:
309 xminusxbars += (i-average)**2
310 if num > 1:
311 sd = math.sqrt(xminusxbars/(num-1))
312 se = sd / math.sqrt(num)
313 if (arguments.marked and locus in marked_set):
314 output.append([gene, average, num, sd, se, "M", "\n"])
315 else:
316 output.append([gene, average, num, sd, se, "\n"])
317 with open(arguments.summary, "wb") as csvfile:
318 writer = csv.writer(csvfile)
319 writer.writerows(output)
320
321
322 #Test: python ../script/aggregate.py -m tigr4_normal.txt -w 1 -x 10 -l 50 -b 0 -c NC_003028b2.gbk -o aggregates/L3_2394eVI_GlucTEST.csv results/L3_2394eVI_Gluc.csv
323
324 #Perl Test: perl ../script/aggregate.pl -m tigr4_normal.txt -w 1 -x 10 -l 50 -b 0 -c NC_003028b2.gbk -o aggregates/L3_2394eVI_Gluc.csv results/L3_2394eVI_Gluc.csv