1 | #!/usr/bin/env python |
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2 | """ |
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3 | run megablast for metagenomics data |
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4 | |
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5 | usage: %prog [options] |
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6 | -d, --db_build=d: The database to use |
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7 | -i, --input=i: Input FASTQ candidate file |
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8 | -w, --word_size=w: Size of best perfect match |
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9 | -c, --identity_cutoff=c: Report hits at or above this identity |
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10 | -e, --eval_cutoff=e: Expectation value cutoff |
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11 | -f, --filter_query=f: Filter out low complexity regions |
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12 | -x, --index_dir=x: Data index directory |
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13 | -o, --output=o: Output file |
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14 | |
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15 | usage: %prog db_build input_file word_size identity_cutoff eval_cutoff filter_query index_dir output_file |
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16 | """ |
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17 | |
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18 | import os, subprocess, sys, tempfile |
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19 | from galaxy import eggs |
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20 | import pkg_resources; pkg_resources.require( "bx-python" ) |
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21 | from bx.cookbook import doc_optparse |
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22 | |
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23 | assert sys.version_info[:2] >= ( 2, 4 ) |
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24 | |
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25 | def stop_err( msg ): |
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26 | sys.stderr.write( "%s\n" % msg ) |
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27 | sys.exit() |
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28 | |
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29 | def __main__(): |
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30 | #Parse Command Line |
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31 | options, args = doc_optparse.parse( __doc__ ) |
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32 | |
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33 | db_build = options.db_build |
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34 | query_filename = options.input.strip() |
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35 | output_filename = options.output.strip() |
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36 | mega_word_size = options.word_size # -W |
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37 | mega_iden_cutoff = options.identity_cutoff # -p |
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38 | mega_evalue_cutoff = options.eval_cutoff # -e |
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39 | mega_temp_output = tempfile.NamedTemporaryFile().name |
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40 | mega_filter = options.filter_query # -F |
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41 | GALAXY_DATA_INDEX_DIR = options.index_dir |
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42 | DB_LOC = "%s/blastdb.loc" % GALAXY_DATA_INDEX_DIR |
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43 | |
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44 | # megablast parameters |
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45 | try: |
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46 | int( mega_word_size ) |
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47 | except: |
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48 | stop_err( 'Invalid value for word size' ) |
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49 | try: |
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50 | float(mega_iden_cutoff) |
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51 | except: |
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52 | stop_err( 'Invalid value for identity cut-off' ) |
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53 | try: |
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54 | float(mega_evalue_cutoff) |
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55 | except: |
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56 | stop_err( 'Invalid value for Expectation value' ) |
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57 | |
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58 | # prepare the database |
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59 | db = {} |
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60 | for i, line in enumerate( file( DB_LOC ) ): |
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61 | line = line.rstrip( '\r\n' ) |
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62 | if not line or line.startswith( '#' ): |
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63 | continue |
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64 | fields = line.split( '\t' ) |
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65 | db[ fields[0] ] = fields[1] |
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66 | |
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67 | if not db.has_key( db_build ): |
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68 | stop_err( 'Cannot locate the target database. Please check your location file.' ) |
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69 | |
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70 | # arguments for megablast |
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71 | chunk = db[ ( db_build ) ] |
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72 | megablast_command = "megablast -d %s -i %s -o %s -m 8 -a 8 -W %s -p %s -e %s -F %s > /dev/null " \ |
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73 | % ( chunk, query_filename, mega_temp_output, mega_word_size, mega_iden_cutoff, mega_evalue_cutoff, mega_filter ) |
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74 | |
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75 | print megablast_command |
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76 | |
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77 | tmp = tempfile.NamedTemporaryFile().name |
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78 | try: |
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79 | tmp_stderr = open( tmp, 'wb' ) |
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80 | proc = subprocess.Popen( args=megablast_command, shell=True, stderr=tmp_stderr.fileno() ) |
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81 | returncode = proc.wait() |
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82 | tmp_stderr.close() |
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83 | # get stderr, allowing for case where it's very large |
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84 | tmp_stderr = open( tmp, 'rb' ) |
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85 | stderr = '' |
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86 | buffsize = 1048576 |
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87 | try: |
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88 | while True: |
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89 | stderr += tmp_stderr.read( buffsize ) |
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90 | if not stderr or len( stderr ) % buffsize != 0: |
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91 | break |
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92 | except OverflowError: |
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93 | pass |
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94 | tmp_stderr.close() |
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95 | if returncode != 0: |
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96 | raise Exception, stderr |
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97 | if os.path.exists( tmp ): |
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98 | os.unlink( tmp ) |
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99 | except Exception, e: |
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100 | if os.path.exists( mega_temp_output ): |
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101 | os.unlink( mega_temp_output ) |
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102 | if os.path.exists( tmp ): |
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103 | os.unlink( tmp ) |
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104 | stop_err( 'Error indexing reference sequence. ' + str( e ) ) |
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105 | |
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106 | output = open( output_filename, 'w' ) |
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107 | invalid_lines = 0 |
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108 | for i, line in enumerate( file( mega_temp_output ) ): |
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109 | line = line.rstrip( '\r\n' ) |
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110 | fields = line.split() |
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111 | try: |
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112 | # get gi and length of that gi seq |
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113 | gi, gi_len = fields[1].split( '_' ) |
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114 | # convert the last column (causing problem in filter tool) to float |
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115 | fields[-1] = float( fields[-1] ) |
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116 | new_line = "%s\t%s\t%s\t%s\t%0.1f" % ( fields[0], gi, gi_len, '\t'.join( fields[2:-1] ), fields[-1] ) |
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117 | except: |
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118 | new_line = line |
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119 | invalid_lines += 1 |
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120 | output.write( "%s\n" % new_line ) |
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121 | output.close() |
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122 | |
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123 | if os.path.exists( mega_temp_output ): |
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124 | os.unlink( mega_temp_output ) #remove the tempfile that we just reformatted the contents of |
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125 | |
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126 | if invalid_lines: |
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127 | print "Unable to parse %d lines. Keep the default format." % invalid_lines |
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128 | |
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129 | # megablast generates a file called error.log, if empty, delete it, if not, show the contents |
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130 | if os.path.exists( './error.log' ): |
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131 | for i, line in enumerate( file( './error.log' ) ): |
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132 | line = line.rstrip( '\r\n' ) |
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133 | print line |
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134 | os.remove( './error.log' ) |
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135 | |
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136 | if __name__ == "__main__" : __main__() |
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