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  • Reciprocal Blast & Parse and Collect best hits

    Hi everyone,

    I'm novice doing reciprocal blast and I have spent a lot of time trying it. I don't know how to do a reciprocal blast and how to parse the best hits and compare & find matches. So, an advice would be a big help to me.

    I need to do a reciprocal Blast in Python (Jupyter) to identify same/ortholougs proteins bewteen two strains.

    - For that, first I make a db with the respective FASTA with the coding sequences.
    Code:
    !makeblastdb -in $fastap_1 -dbtype 'prot'
    !makeblastdb -in $fastap_2 -dbtype 'prot'
    - Then, I run blastp for both files:
    Code:
    blastp_ab = !blastp -query $fastap_1 -db $fastap_2 -out ab.txt
    blastp_ba = !blastp -query $fastap_2 -db $fastap_1 -out ba.txt
    Up to here everything works. **Now, I have to parse the blast results to collect best hits. I have tried it,** **but not even print "hello??"**

    If I pass as argument the file "ab.txt", the function "collect_best_hits" returns me an empty dictionary. And if I pass the handle "blastp_ab" as argument, "collect_best_hits" functions return me and advice of "blastall" and the empty dictionary:

    > no BLAST output. Check that blastall is in your PATH
    >
    > { }


    (Options)
    Code:
    #filename = "ab.txt"
    #filename = blastp_ab
    filename = open("ab.txt", "r")
    
    collect_best_hits(filename)

    The function contains:

    Code:
    import os
    import sys
    import csv
    import blastparser
    
    
    def collect_best_hits(filename):
           d = {}
           try:
                for n, record in enumerate(blastparser.parse_fp(filename)):
                    print "hola"
                    if n % 100 == 0:
                          print >>sys.stderr, 'loading 1 ...', n
                   
                    print "!-", n, record
        
                    best_score = None
                    for hit in record.hits:
                        print "!- ", hit, len(record.hits)
                        for match in hit.matches:
                            print "!- ", match, len(hit.matches)
                            query = record.query_name
                            if query.startswith('gi'):
                                query = query.split('|', 2)[2]
                            subject = hit.subject_name
        
                            score = match.score
                            print "!- ", score
        
                            # only keep the best set of scores for any query
                            if best_score and best_score > score:
                                continue
                            best_score = score
        
                            x = d.get(query, [])
                            x.append((subject, score))
                            d[query] = x
        
                        if best_score and best_score != score:
                            break
            except Exception as e:
                print(e)
            return d

    What I'm doing wrong?

    All help is welcome!

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