Harvard:Biophysics 101/2007/Notebook:Michael Wang/2007-3-6

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(New page: Notes: This isn't as clean as the sample output, but it compares two hardcoded strings and does mutation compatison. The functions package mutations and orfs into objects that could be ex...)
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<pre>
<pre>
#find all orfs within the sequences
#find all orfs within the sequences
-
import re
+
import re, string
from Bio import Transcribe
from Bio import Transcribe
transcriber = Transcribe.unambiguous_transcriber
transcriber = Transcribe.unambiguous_transcriber
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teststring = re.sub("\s+", "", teststring)
teststring = re.sub("\s+", "", teststring)
teststring2 = re.sub("\s+", "", teststring2)
teststring2 = re.sub("\s+", "", teststring2)
-
print "Forward"
+
print "Forward orfs"
allorfs = findORFS(teststring)
allorfs = findORFS(teststring)
allorfs2 = findORFS(teststring2)
allorfs2 = findORFS(teststring2)
temp_file = open(os.path.join(os.curdir, 'temp.txt'),"w")
temp_file = open(os.path.join(os.curdir, 'temp.txt'),"w")
-
temp_file.write(">temp1 ")
+
temp_file.write(">temp1|\n ")
temp_file.write(teststring)
temp_file.write(teststring)
temp_file.write("\n\n")
temp_file.write("\n\n")
-
temp_file.write(">gtemp2 ")
+
temp_file.write(">gtemp2|\n ")
temp_file.write(teststring2)
temp_file.write(teststring2)
temp_file.close()
temp_file.close()
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reverse_list1.reverse()
reverse_list1.reverse()
reverse_list2.reverse()
reverse_list2.reverse()
-
 
reverse_string1 = "".join(reverse_list1)
reverse_string1 = "".join(reverse_list1)
reverse_string2 = "".join(reverse_list2)
reverse_string2 = "".join(reverse_list2)
-
print "reverse"
+
table = string.maketrans("ATGC","TACG")
 +
reverse_string1 = reverse_string1.translate(table)
 +
reverse_string2 = reverse_string2.translate(table)
 +
print "reverse orfs"
revallorfs = findORFS(reverse_string1)
revallorfs = findORFS(reverse_string1)
revallorfs2 = findORFS(reverse_string2)
revallorfs2 = findORFS(reverse_string2)
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 +
 +
 +
 +
</pre>
</pre>

Revision as of 01:31, 6 March 2007

Notes: This isn't as clean as the sample output, but it compares two hardcoded strings and does mutation compatison. The functions package mutations and orfs into objects that could be exported for further manipulation. Though this version doesn't implement it, it would be fairly easy to load in multiple sequences to do pairwise alignments and call the main script (This also goes for the reverse strand comparisons). Also missing in this version is the final output proteins for the insertions and deletions. I couldn't quite figure out exactly we're supposed to handle out of fram mutations in getting the codons to line up. This also generates an error code from clustalw though the output seems to be fine.

#find all orfs within the sequences
import re, string
from Bio import Transcribe
transcriber = Transcribe.unambiguous_transcriber
from Bio import Translate
from Bio.Alphabet import IUPAC
from Bio.Seq import Seq
import os
from Bio import Clustalw
standard_translator = Translate.unambiguous_dna_by_id[1]

class codon:
    def  __init__(self, sequence=""):
        self.sequence = sequence
        self.mRNA = transcriber.transcribe(Seq(sequence, IUPAC.unambiguous_dna))
        self.protein = standard_translator.translate(Seq(sequence, IUPAC.unambiguous_dna))[0]

class raw_mutation:
    def __init__(self,type="",ref_location=0,other_location=0, original="", mutant=""):
        self.type = type
        self.ref_location=ref_location
        self.other_location = other_location
        self.original = original
        self.mutant = mutant

class mutation:
    def __init__(self,type="",ref_span=[0,0],other_span=[0,0],original="", mutant=""):
        self.type = type
        self.ref_span = ref_span
        self.other_span = other_span
        self.original = original
        self.mutant = mutant
    
class orf:
    #On initiation, the orf is stored as a list of codons.  If the orf has no stop, any excess bases will
    #be ignored on the conversion to codons
    def __init__(self, source="", source_span=[]):
        self.source = source
        self.codons = []
        self.sequence = source[source_span[0]:source_span[1]]
        self.source_span = source_span
        for i in range(len(self.sequence)/3):
            temp_codon = codon(self.sequence[i*3:3+(i*3)])  #this algorithm of seperating into codons ignores any excess bases
            self.codons.append(temp_codon)
        
    #orfs are indexed by codons
    def __getitem__(self,index):
        return self.codons[index]

def find_mutations(self,other):
    refPos = 0
    seqPos = 0
    #print "Ref:",self
    #print "Other:",other
    all_mutations = []
    for i in range(len(self)):
        #print self[i], " ", other[i], ' ', refPos, ' ', seqPos
        if self[i] != other[i]:
            if self[i] == '-':
                new_mutation = raw_mutation("insertion",refPos,seqPos)
            elif other[i] == '-':
                new_mutation = raw_mutation("deletion",refPos,seqPos)
            else:
                new_mutation = raw_mutation("point",refPos,seqPos, self[i],other[i])
            all_mutations.append(new_mutation)
            #print "Mutation!"
            #print new_mutation.type," ",new_mutation.ref_location," ",new_mutation.other_location
             
        if self[i] != '-':
            refPos += 1
        if other[i] != '-':
            seqPos += 1
    return all_mutations
            
def consolidate_mutations(raw_mutations):
    #I think this will have a bug for insertions before the first base of the refseq because
    #in every other case, ref_loc will refer to the base before the insertion
    consolidated_mutations =[]
    i = 0
    while i < len(raw_mutations):
        #print "outer loop"
        current_type = raw_mutations[i].type
        if(current_type == "point"):
            ref_loc = raw_mutations[i].ref_location
            other_loc = raw_mutations[i].other_location
            new_mutation = mutation("point",[ref_loc,ref_loc],[other_loc,other_loc],raw_mutations[i].original, raw_mutations[i].mutant)
            consolidated_mutations.append(new_mutation)
            i += 1
        elif(current_type == "deletion") or (current_type == "insertion"):
            
            ref_start = raw_mutations[i].ref_location
            other_start = raw_mutations[i].other_location
            ref_end = raw_mutations[i].ref_location
            other_end = raw_mutations[i].other_location 
            i += 1
            #It would have been nice to have a do while here...
            while (i<=len(raw_mutations)): #and (raw_mutations[i].type == current_type):
                #hopefully it exits after the first condition so it doesn't go past array length
                if (i==len(raw_mutations)) or ((raw_mutations[i].ref_location!=ref_start) and (raw_mutations[i].other_location!=other_start)):
                    new_mutation = mutation(current_type,[ref_start,ref_end],[other_start,other_end])
                    consolidated_mutations.append(new_mutation)
                    #print current_type, "found at ", "ref", ref_start, ",",ref_end," seq ", other_start, ",", other_end
                    break
                elif (current_type == "insertion"): #and (raw_mutations[i].ref_location==ref_start):
                    other_end += 1
                    i += 1
                    #print "extending insertion"
                elif (current_type == "deletion"): #and (raw_mutations[i].ref_location==other_start):
                    ref_end += 1
                    i += 1
                    #print "extending deletion"
                else:
                    print "I shouldn't be here!!!"
                    break

    return consolidated_mutations
        
def findORFS(sequence, startpos=0):
    print "For",sequence
    all_orfs = []
    start = re.compile('ATG')
    stop = re.compile('(TAA|TGA|TAG)')
    all_starts = start.finditer(sequence)
    all_stops = stop.finditer(sequence)
    all_starts_list = []
    for match in all_starts:
        all_starts_list.append(match.span())
    all_stops_list = []
    for stops in all_stops:
        all_stops_list.append(stops.span())
        #print stops.span()
    for start in all_starts_list:
        found = 0
        for stop in all_stops_list:
            diff = (stop[0]-start[0])
            if ((diff>0) and ((diff%3) == 0)):
                print "orf at:", start[0]," ",stop[1]
                temp_orf = orf(sequence,(start[0],stop[1]))
                all_orfs.append(temp_orf)
                #all_orfs.append((start[0],stop[1]))
                found = 1
                break
        if found ==0:
            print "open orf",
            temp_orf = orf(sequence,(start[0],None))
            all_orfs.append(temp_orf)
            print "starting at:", start[0]
            #all_orfs.append((start[0],None))
        
    return all_orfs

def check_point(mutation, ref_orfs):
    mut_pos = mutation.ref_span[0] #ref_span[0] and [1] are the same
    print "For ", mutation.type, "at position", mutation.ref_span[0], ':'
    for i in ref_orfs:
        if (mut_pos>=i.source_span[0] and mut_pos<=i.source_span[1]): #does the mutation affect the orf
            print "Point affects ", i.sequence
            ref_codon_position = (mut_pos - i.source_span[0])/3
            
            base_position = (mut_pos-i.source_span[0])%3
            original_aa = i[ref_codon_position].protein
            mut_codon = i[ref_codon_position].sequence
         #   print "Original Codon:", mut_codon
            mut_codon = list(mut_codon)
            mut_codon[base_position] = mutation.mutant
            mut_codon = "".join(mut_codon)
         #   print "Mutant Codon",mut_codon
            mutant_aa = standard_translator.translate(Seq(mut_codon, IUPAC.unambiguous_dna))[0]
            if (mutant_aa == original_aa):
                print "Silent Point Mutation detected: ", 
            else:
                print "Non-silent Point Mutation detected", 
            print (mutation.original+str(mut_pos)+mutation.mutant),"Protein result:",original_aa+str(mut_pos)+mutant_aa

def formatted_seq_print (sequence):
    for i in range(len(sequence)/3):
        start_base = i*3
        print_codon = sequence.data[start_base:start_base+3]
        print print_codon, 
    print ""

def formatted_codon_print (sequence):
    for i in range(len(sequence)/3):
        start_base = i*3
        prot = standard_translator.translate(Seq(sequence, IUPAC.unambiguous_dna))[0]
        print_codon = sequence.data[start_base:start_base+3]
        #if currently in the deletion region, don't print
        print print_codon, 
    print ""
        
        

def check_frame(mutation, all_orfs, all_records):
    if ((mutation.ref_span[1]-mutation.ref_span[0])%3 !=0) or ((mutation.other_span[1]-mutation.other_span[0])%3 !=0):
        print mutation.type, "from", mutation.ref_span[0]," to", mutation.ref_span[1], "is a frameshift"
    else:
        print mutation.type, "from", mutation.ref_span[0]," to", mutation.ref_span[1], "is in frame"
    
    print "odna:",
    formatted_seq_print (all_records[0].seq)
    #for i in range(len(something)/3):
    #    print " ",some_protein," "
    print "mdna:",
    formatted_seq_print (all_records[1].seq)
    #for i in range(len(something)/3):
    #    print " ",some_protein," "

        

#Main Program starts here          
teststring = "  A TGG GGG GGA ATG ATT AAC GTC GTT AAA GTA TGT TTT TT"
teststring2= "CGA ATG GGG GCG ATG ATT AAC C GTT AAA GTA TGT TTT TTG TAG"
print teststring
teststring = re.sub("\s+", "", teststring)
teststring2 = re.sub("\s+", "", teststring2)
print "Forward orfs"
allorfs = findORFS(teststring)
allorfs2 = findORFS(teststring2)
temp_file = open(os.path.join(os.curdir, 'temp.txt'),"w")
temp_file.write(">temp1|\n ")
temp_file.write(teststring)
temp_file.write("\n\n")
temp_file.write(">gtemp2|\n ")
temp_file.write(teststring2)
temp_file.close()
cline = Clustalw.MultipleAlignCL(os.path.join(os.curdir, 'temp.txt'))
cline.set_output('test.aln')
alignment = Clustalw.do_alignment(cline)
all_records = alignment.get_all_seqs()
#print alignment
mutations = find_mutations(all_records[0].seq,all_records[1].seq)
#print mutations
all_real = consolidate_mutations (mutations)
for i in all_real:
    print i.type, "at ref:", i.ref_span, " other ", i.other_span
for i in all_real:
    if (i.type == "point"):
        check_point(i, allorfs)
    else:
        check_frame(i,allorfs,all_records)



reverse_list1 = list(teststring)[:]
reverse_list2 = list(teststring2)[:]
teststring = "".join(teststring)
teststring2 = "".join(teststring2)
reverse_list1.reverse()
reverse_list2.reverse()
reverse_string1 = "".join(reverse_list1)
reverse_string2 = "".join(reverse_list2)
table = string.maketrans("ATGC","TACG")
reverse_string1 = reverse_string1.translate(table)
reverse_string2 = reverse_string2.translate(table)
print "reverse orfs"
revallorfs = findORFS(reverse_string1)
revallorfs2 = findORFS(reverse_string2)
#print "orfs2"
#for i in allorfs2:
#    print i.sequence
        
        





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