This file is indexed.

/usr/bin/bp_classify_hits_kingdom is in bioperl 1.6.924-1.

This file is owned by root:root, with mode 0o755.

The actual contents of the file can be viewed below.

  1
  2
  3
  4
  5
  6
  7
  8
  9
 10
 11
 12
 13
 14
 15
 16
 17
 18
 19
 20
 21
 22
 23
 24
 25
 26
 27
 28
 29
 30
 31
 32
 33
 34
 35
 36
 37
 38
 39
 40
 41
 42
 43
 44
 45
 46
 47
 48
 49
 50
 51
 52
 53
 54
 55
 56
 57
 58
 59
 60
 61
 62
 63
 64
 65
 66
 67
 68
 69
 70
 71
 72
 73
 74
 75
 76
 77
 78
 79
 80
 81
 82
 83
 84
 85
 86
 87
 88
 89
 90
 91
 92
 93
 94
 95
 96
 97
 98
 99
100
101
102
103
104
105
106
107
108
109
110
111
112
113
114
115
116
117
118
119
120
121
122
123
124
125
126
127
128
129
130
131
132
133
134
135
136
137
138
139
140
141
142
143
144
145
146
147
148
149
150
151
152
153
154
155
156
157
158
159
160
161
162
163
164
165
166
167
168
169
170
171
172
173
174
175
176
177
178
179
180
181
182
183
184
185
186
187
188
189
190
191
192
193
194
195
196
197
198
199
200
201
202
203
204
205
206
207
208
209
210
211
212
213
214
215
216
217
218
219
220
221
222
223
224
225
#!/usr/bin/perl 

eval 'exec /usr/bin/perl  -S $0 ${1+"$@"}'
    if 0; # not running under some shell

=head1 NAME

bp_classify_hits_kingdom - classify BLAST hits by taxonomic kingdom

=head2 USAGE

bp_classify_hits_kingdom [-i tab_file] [-i second_BLAST_file] [-e evalue_cutoff]
                      [-t dir_where_TAXONOMY_files_are] [-g gi2taxid] 
                      [-z PATH_TO_zcat] [-v]

=head2 DESCRIPTION

Will print out the taxonomic distribution (at the kingdom level) for a
set of hits against the NR database.  By default, this script assumes you
did a search against the protein database (gi_taxid_nuc.dump file).

This expects BLAST files in tabbed -m9 or -m8 format.  Output with -m
8 or use blast2table.pl to convert (or fastam9_to_table.PLS if using
FASTA).

  Input values:
    -t/--taxonomy Directory where the taxonomy .dmp files are (from NCBI)
    -g/--gi       File path of the gi2taxid file (gi_taxid_prot.dmp for proteins
                  or gi_taxid_nucl.dmp if the search was against a nucleid database)
    -i/--in       The name of the tab delimited -m8/-m9 output files to process
    -e/--evalue   Provide an E-value cutoff for hits to be considered
    -z/--zcat     Path to the 'zcat' executable, can also be 'gunzip -c'
                  if no zcat on your system.
  Flags:
    -v/--verbose  To turn on verbose messages
    -h/--help     Display this helpful information

This is intended to be useful starting script, but users may want to
customize the output and parameters. Note that I am summarizing the
kingdoms here and Eukaryota not falling into Metazoa, Viridiplantae, or
Fungi gets grouped into the general superkingdom Eukaryota for simplicity.
There are comments in the code directing you to where changes can be made
if you wanted to display hits by phylum for example.  Note that you must
wipe out the cache file 'gi2class' that is created in your directory after
making these changes.

=head2 AUTHOR

Jason Stajich jason_at_bioperl_dot_org

=cut

use strict;
use warnings;
use Bio::DB::Taxonomy;
use DBI;
use Env;
use File::Spec;
use vars qw($SEP);
my $DEBUG = 0;
use Getopt::Long;
$SEP = '_';

my $evalue_filter = 1e-3;
my @files;
my $zcat = 'zcat'; # or gunzip -c 
my $prefix = File::Spec->catfile($HOME,'taxonomy');
my $gi2taxidfile = "$prefix/gi_taxid_prot.dmp.gz";
my $force = 0; # don't use the cached gi2taxid file
GetOptions(
       'v|verbose|debug' => \$DEBUG,
       'force!'          => \$force,
       'z|zcat:s'        => \$zcat,
       'i|in:s'          => \@files,
       'e|evalue:f'      => \$evalue_filter,
       't|taxonomy:s'    => \$prefix,
       'g|gi|gi2taxid:s' => \$gi2taxidfile,
       'h|help'          => sub { system('perldoc', $0); exit },
       );

# insure idx location is created
mkdir(File::Spec->catfile($prefix,'idx')) 
    unless -d File::Spec->catfile($prefix,'idx');

# these files came from ftp://ftp.ncbi.nih.gov/pub/taxonomy
my $taxdb = Bio::DB::Taxonomy->new
    (-source => 'flatfile',
     -directory => File::Spec->catfile
     ($prefix, 'idx'), 
     -nodesfile => File::Spec->catfile($prefix,'nodes.dmp'),
     -namesfile => File::Spec->catfile($prefix,'names.dmp')
     );
my %query;

my (%taxid4gi,%gi2node);
my $dbh = tie(%gi2node, 'DB_File', 'gi2class');
my $giidxfile = File::Spec->catfile($prefix,'idx','gi2taxid');
my $done = -f $giidxfile;
$done = 0 if $force;
my $dbh2 = $dbh = DBI->connect("dbi:SQLite:dbname=$giidxfile","","");
if( ! $done ) {
    $dbh2->do("CREATE TABLE gi2taxid ( gi integer PRIMARY KEY,
                      taxid integer NOT NULL)");
    $dbh2->{AutoCommit} = 0;
    my $fh;
    # this file came from ftp://ftp.ncbi.nih.gov/pub/taxonomy
    # I'm interested in protein hits therefor _prot file.
    if (not -f $gi2taxidfile) {
        die "Error: File $gi2taxidfile does not exist\n";
    }
    if( $gi2taxidfile =~ /\.gz$/ ) {
        open($fh, "$zcat $gi2taxidfile |" ) || die "$zcat $gi2taxidfile: $!";
    } else {
        open($fh, $gi2taxidfile ) || die "Error: could not read file $gi2taxidfile: $!";
    }
    my $i = 0;
    my $sth = $dbh2->prepare("INSERT INTO gi2taxid (gi,taxid) VALUES (?,?)");

    while(<$fh>) {
        my ($gi,$taxid) = split;
        $sth->execute($gi,$taxid);
        $i++;
        if( $i % 500000 == 0 ) {
            $dbh->commit;
            warn("$i\n") if $DEBUG;
        } 
    }
    $dbh->commit;
    $sth->finish;
}

for my $file ( @files ) {
    warn("$file\n");
    my $gz;
    if( $file =~ /\.gz$/) {
        $gz = 1;
    }
    my ($spname) = split(/\./,$file); 
    my ($fh,$i);
    if( $gz ) {
        open($fh, "$zcat $file |")  || die "$zcat $file: $!";
    } else {
        open($fh, $file) || die "$file: $!";
    }
    my $sth = $dbh->prepare("SELECT taxid from gi2taxid WHERE gi=?");
    while(<$fh>) {
        next if /^\#/;
        my ($qname,$hname,$pid,$qaln,$mismatch,$gaps,
            $qstart,$qend,$hstart,$hend,
            $evalue,$bits,$score) = split(/\t/,$_);        
        next if( $evalue > $evalue_filter );
        if( ! exists $query{$spname}->{$qname} ) {
            $query{$spname}->{$qname} = {};
        }
        
        if( $hname =~ /gi\|(\d+)/) {
            my $gi = $1;
            if( ! $gi2node{$gi} ){ # see if we cached the results from before
                $sth->execute($gi);
                my $taxid;
                $sth->bind_columns(\$taxid);
                if( ! $sth->fetch ) {
                    warn("no taxid for $gi\n");
                    next;
                }
                my $node = $taxdb->get_Taxonomy_Node($taxid);
                if( ! $node ) {
                    warn("cannot find node for gi=$gi ($hname) (taxid=$taxid)\n");
                    next;
                }
                my $parent = $taxdb->get_Taxonomy_Node($node->parent_id);

                # THIS IS WHERE THE KINGDOM DECISION IS MADE
                # DON'T FORGET TO WIPE OUT YOUR CACHE FILE
                # gi2class after you make changes here
                while( defined $parent && $parent->node_name ne 'root' ) { 
                    # this is walking up the taxonomy hierarchy
                    # can be a little slow, but works...
                    #warn( "\t",$parent->rank, " ", $parent->node_name, "\n");
                    # deal with Eubacteria, Archea separate from 
                    # Metazoa, Fungi, Viriplantae differently
                    # (everything else Eukaryotic goes in Eukaryota)
                    if( $parent->rank eq 'kingdom') {
                        # caching in ... 
                        ($gi2node{$gi}) = $parent->node_name;
                        last;
                    } elsif( $parent->rank eq 'superkingdom' ) {
                        # caching in ... 
                        ($gi2node{$gi}) = $parent->node_name;
                        $gi2node{$gi} =~ s/ \<(bacteria|archaea)\>//g;
                        last;
                    }
                    $parent = $taxdb->get_Taxonomy_Node($parent->parent_id);
                }
            } 
            my ($kingdom) = $gi2node{$gi};
            #warn("$gi2node{$gi}\n");
            unless( defined $kingdom && length($kingdom) ) {
                #warn("no kingdom for $hname\n");
            } else {
                $query{$spname}->{$qname}->{$kingdom}++;
            }
        } else {
            warn("no GI in $hname\n");
        }
    }
    last if ( $DEBUG && $i++ > 10000);
    $sth->finish;
}

# print out the taxonomic distribution
while( my ($sp,$d) = each %query ) {
    my $total = scalar keys %$d;
    print "$sp total=$total\n";
    my %seen;
    for my $v ( values %$d ) {
        my $tag = join(",",sort keys %$v );
        $seen{$tag}++;
    }
    for my $t ( sort { $seen{$a} <=> $seen{$b} } keys %seen ) {
        printf " %-20s\t%d\t%.2f%%\n",
        $t,$seen{$t}, 100 * $seen{$t} / $total;
    }
    print "\n\n";
}