| 1 | #!/usr/bin/perl |
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| 2 | |
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| 3 | ############################################################################# |
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| 4 | # # |
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| 5 | # File: arb_sativa.pl # |
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| 6 | # Purpose: Adapter script for SATIVA taxonomy validation pipeline # |
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| 7 | # # |
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| 8 | # Author: Alexey Kozlov (alexey.kozlov@h-its.org) # |
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| 9 | # 2014 HITS gGmbH / Exelixis Lab # |
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| 10 | # http://h-its.org http://www.exelixis-lab.org/ # |
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| 11 | # # |
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| 12 | ############################################################################# |
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| 13 | |
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| 14 | use strict; |
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| 15 | use warnings; |
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| 16 | # use diagnostics; |
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| 17 | |
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| 18 | BEGIN { |
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| 19 | if (not exists $ENV{'ARBHOME'}) { die "Environment variable \$ARBHOME has to be defined"; } |
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| 20 | my $arbhome = $ENV{'ARBHOME'}; |
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| 21 | push @INC, "$arbhome/lib"; |
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| 22 | push @INC, "$arbhome/PERL_SCRIPTS/lib"; |
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| 23 | 1; |
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| 24 | } |
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| 25 | |
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| 26 | use ARB; |
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| 27 | use tools; |
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| 28 | |
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| 29 | my $sativa_home = $ENV{'ARBHOME'}.'/bin/sativa'; |
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| 30 | my ($start_time, $trainer_time, $mislabels_time); |
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| 31 | |
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| 32 | # ------------------------------------------------------------ |
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| 33 | |
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| 34 | sub expectEntry($$) { |
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| 35 | my ($gb_father,$key) = @_; |
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| 36 | my $gb = ARB::search($gb_father, $key, 'NONE'); |
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| 37 | defined $gb || die "Expected entry '$key' in '".ARB::get_db_path($gb_father)."'"; |
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| 38 | return $gb; |
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| 39 | } |
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| 40 | |
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| 41 | # ----------------------- |
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| 42 | |
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| 43 | sub exportTaxonomy($$$$$\@) { |
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| 44 | my ($seq_file,$tax_file,$tax_field,$sp_field,$marked_only,@marklist_r) = @_; |
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| 45 | |
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| 46 | open(TAXFILE,'>'.$tax_file) || die "can't open '$tax_file' (Reason: $!)"; |
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| 47 | # open(SEQFILE,'>'.$seq_file) || die "can't open '$seq_file' (Reason: $!)"; |
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| 48 | |
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| 49 | my $gb_main = ARB::open(":","r"); |
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| 50 | $gb_main || expectError('db connect (no running ARB?)'); |
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| 51 | |
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| 52 | dieOnError(ARB::begin_transaction($gb_main), 'begin_transaction'); |
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| 53 | |
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| 54 | for (my $gb_species = BIO::first_species($gb_main); |
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| 55 | $gb_species; |
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| 56 | $gb_species = BIO::next_species($gb_species)) |
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| 57 | { |
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| 58 | my $marked = ARB::read_flag($gb_species); |
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| 59 | if ($marked==1 || $marked_only==0) { |
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| 60 | my $acc_no = BIO::read_string($gb_species, "name"); |
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| 61 | $acc_no || expectError('read_string acc'); |
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| 62 | my $tax = BIO::read_string($gb_species, $tax_field); |
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| 63 | $tax || expectError('read_string '.$tax_field); |
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| 64 | my $species_name = BIO::read_string($gb_species, $sp_field); |
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| 65 | my $lineage = $tax; |
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| 66 | if (substr($lineage, -1) eq ';') { |
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| 67 | chop($lineage); |
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| 68 | } |
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| 69 | if (defined $species_name) { |
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| 70 | $lineage = $lineage.";".$species_name; |
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| 71 | } |
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| 72 | |
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| 73 | print TAXFILE $acc_no."\t".$lineage."\n"; |
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| 74 | |
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| 75 | # my $gb_ali = expectEntry($gb_species, 'ali_16s'); |
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| 76 | # my $gb_data = expectEntry($gb_ali, 'data'); |
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| 77 | # my $seq = ARB::read_string($gb_data); |
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| 78 | |
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| 79 | # print SEQFILE ">".$acc_no."\n"; |
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| 80 | # print SEQFILE $seq."\n"; |
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| 81 | |
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| 82 | push(@marklist_r, $acc_no); |
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| 83 | } |
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| 84 | } |
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| 85 | |
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| 86 | ARB::commit_transaction($gb_main); |
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| 87 | ARB::close($gb_main); |
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| 88 | |
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| 89 | close(TAXFILE); |
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| 90 | # close(SEQFILE); |
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| 91 | } |
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| 92 | |
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| 93 | # ------------------------------------------------------------ |
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| 94 | |
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| 95 | sub cpu_has_feature($) { |
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| 96 | my ($feature) = @_; |
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| 97 | my $cmd; |
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| 98 | my $uname = `uname`; |
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| 99 | chomp($uname); |
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| 100 | |
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| 101 | if ($uname eq "Darwin") { |
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| 102 | $cmd = "sysctl machdep.cpu.features"; |
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| 103 | } |
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| 104 | elsif ($uname eq "Linux") { |
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| 105 | $cmd = "grep flags /proc/cpuinfo"; |
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| 106 | } |
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| 107 | else { |
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| 108 | return 0; |
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| 109 | } |
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| 110 | |
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| 111 | `$cmd | grep -qi "$feature"`; |
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| 112 | return $? + 1; |
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| 113 | } |
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| 114 | |
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| 115 | sub cpu_get_cores() { |
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| 116 | my $cores = 2; |
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| 117 | my $uname = `uname`; |
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| 118 | chomp($uname); |
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| 119 | if ($uname eq "Darwin") { |
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| 120 | $cores = `sysctl -n hw.ncpu`; |
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| 121 | } |
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| 122 | elsif ($uname eq "Linux") { |
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| 123 | $cores = `grep -c "^processor" /proc/cpuinfo`; |
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| 124 | } |
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| 125 | |
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| 126 | return $cores; |
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| 127 | } |
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| 128 | |
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| 129 | sub runPythonPipeline($$$$$) { |
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| 130 | my ($seq_file,$tax_file,$dup_rank_names,$wrong_rank_count,$rank_test) = @_; |
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| 131 | |
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| 132 | my $refjson_file = 'arb_export.json'; |
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| 133 | my $cfg_file = $sativa_home.'/epac.cfg.sativa'; |
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| 134 | |
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| 135 | # auto-configure RAxML |
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| 136 | my $cores = cpu_get_cores(); |
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| 137 | |
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| 138 | my $bindir = $ENV{'ARBHOME'}."/bin"; |
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| 139 | my $tmpdir = `pwd`; |
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| 140 | chomp($tmpdir); |
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| 141 | |
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| 142 | my $stime = time; |
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| 143 | |
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| 144 | my $trainer_cmd = $sativa_home.'/epa_trainer.py'; |
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| 145 | system($trainer_cmd, '-t', $tax_file, '-s', $seq_file, '-r', $refjson_file, '-c', $cfg_file, '-T', $cores, '-no-hmmer', |
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| 146 | '-dup-rank-names', $dup_rank_names, '-wrong-rank-count', $wrong_rank_count, '-tmpdir', $tmpdir, '-C'); |
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| 147 | |
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| 148 | $trainer_time = time - $stime; |
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| 149 | |
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| 150 | if (-e $refjson_file) { |
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| 151 | $stime = time; |
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| 152 | my $checker_cmd = $sativa_home.'/find_mislabels.py'; |
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| 153 | my @args = ($checker_cmd, '-r', $refjson_file, '-c', $cfg_file, '-T', $cores, '-v', '-tmpdir', $tmpdir); |
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| 154 | if ($rank_test == 1) { push(@args, "-ranktest"); } |
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| 155 | system(@args); |
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| 156 | $mislabels_time = time - $stime; |
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| 157 | } |
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| 158 | else { |
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| 159 | die "There was a problem running SATIVA (see messages above). Exiting..." |
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| 160 | } |
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| 161 | } |
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| 162 | |
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| 163 | # ------------------------------------------------------------ |
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| 164 | |
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| 165 | sub deleteIfExists($$) { |
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| 166 | my ($father,$key) = @_; |
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| 167 | |
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| 168 | my $gb_field = ARB::entry($father,$key); |
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| 169 | if (defined $gb_field) { |
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| 170 | my $error = ARB::delete($gb_field); |
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| 171 | if ($error) { die $error; } |
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| 172 | } |
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| 173 | } |
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| 174 | |
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| 175 | sub importResults($$$) { |
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| 176 | my ($res_file,$marked_only,$mark_misplaced) = @_; |
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| 177 | |
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| 178 | open(FILE,'<'.$res_file) || die "can't open '$res_file' (Reason: $!)"; |
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| 179 | |
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| 180 | my %mis_map = (); |
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| 181 | |
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| 182 | # skip the header |
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| 183 | my $header = <FILE>; |
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| 184 | |
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| 185 | while (my $line = <FILE>) { |
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| 186 | chomp $line; |
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| 187 | my @fields = split "\t" , $line; |
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| 188 | my $seq_id = $fields[0]; |
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| 189 | my $lvl = $fields[1]; |
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| 190 | my $conf = $fields[4]; |
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| 191 | my $new_tax = $fields[6]; |
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| 192 | $mis_map{$seq_id} = {conf => $conf, new_tax => $new_tax, mis_lvl => $lvl}; |
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| 193 | if (scalar(@fields) == 9) { $mis_map{$seq_id}{'rank_conf'} = $fields[8]; } |
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| 194 | } |
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| 195 | close(FILE); |
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| 196 | |
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| 197 | my $gb_main = ARB::open(":","rw"); |
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| 198 | $gb_main || expectError('db connect (no running ARB?)'); |
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| 199 | |
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| 200 | dieOnError(ARB::begin_transaction($gb_main), 'begin_transaction'); |
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| 201 | |
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| 202 | my $count = 0; |
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| 203 | for (my $gb_species = BIO::first_species($gb_main); |
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| 204 | $gb_species; |
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| 205 | $gb_species = BIO::next_species($gb_species)) |
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| 206 | { |
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| 207 | my $error; |
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| 208 | my $name = BIO::read_string($gb_species, "name"); |
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| 209 | $name || expectError('read_string name'); |
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| 210 | my $marked = ARB::read_flag($gb_species); |
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| 211 | if (defined $mis_map{$name}) { |
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| 212 | $error = BIO::write_int($gb_species, "sativa_mislabel_flag", 1); |
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| 213 | if ($error) { die $error; } |
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| 214 | $error = BIO::write_string($gb_species, "sativa_tax", $mis_map{$name}{'new_tax'}); |
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| 215 | if ($error) { die $error; } |
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| 216 | $error = BIO::write_float($gb_species, "sativa_seqmis_conf", $mis_map{$name}{'conf'}); |
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| 217 | if ($error) { die $error; } |
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| 218 | $error = BIO::write_string($gb_species, "sativa_mislabel_level", $mis_map{$name}{'mis_lvl'}); |
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| 219 | if ($error) { die $error; } |
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| 220 | if (exists $mis_map{$name}{'rank_conf'}) { |
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| 221 | $error = BIO::write_string($gb_species, "sativa_rankmis_conf", $mis_map{$name}{'rank_conf'}); |
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| 222 | if ($error) { die $error; } |
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| 223 | } |
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| 224 | |
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| 225 | if ($mark_misplaced==1) { ARB::write_flag($gb_species, 1); } |
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| 226 | $count++; |
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| 227 | } |
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| 228 | elsif ($marked==1 || $marked_only==0) { |
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| 229 | $error = BIO::write_int($gb_species, "sativa_mislabel_flag", 0); |
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| 230 | if ($error) { die $error; } |
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| 231 | |
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| 232 | # deleteIfExists($gb_species, "sativa_tax"); |
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| 233 | # deleteIfExists($gb_species, "sativa_conf"); |
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| 234 | # deleteIfExists($gb_species, "sativa_seqmis_conf"); |
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| 235 | # deleteIfExists($gb_species, "sativa_mislabel_level"); |
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| 236 | # deleteIfExists($gb_species, "sativa_misrank_conf"); |
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| 237 | |
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| 238 | if ($mark_misplaced==1) { ARB::write_flag($gb_species, 0); } |
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| 239 | } |
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| 240 | } |
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| 241 | |
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| 242 | print "\nMislabels found: $count\n\n"; |
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| 243 | ARB::commit_transaction($gb_main); |
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| 244 | ARB::close($gb_main); |
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| 245 | } |
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| 246 | |
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| 247 | |
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| 248 | # ------------------------------------------------------------ |
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| 249 | |
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| 250 | sub die_usage($) { |
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| 251 | my ($err) = @_; |
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| 252 | print "Purpose: Run SATIVA taxonomy validation pipeline\n"; |
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| 253 | print "and import results back into ARB\n"; |
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| 254 | print "Usage: arb_sativa.pl [--marked-only] [--mark-misplaced] [--rank-test] tax_field [species_field]\n"; |
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| 255 | print " tax_field Field contatining full (original) taxonomic path (lineage)\n"; |
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| 256 | print " species_field Field containing species name\n"; |
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| 257 | print " --marked-only Process only species that are marked in ARB (default: process all)\n"; |
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| 258 | print " --mark-misplaced Mark putatively misplaced species in ARB upon completion\n"; |
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| 259 | print " --rank-test Test for misplaced higher ranks\n"; |
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| 260 | print "\n"; |
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| 261 | die "Error: $err\n"; |
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| 262 | } |
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| 263 | |
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| 264 | sub main() { |
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| 265 | my $args = scalar(@ARGV); |
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| 266 | if ($args<1) { die_usage('Missing arguments'); } |
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| 267 | |
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| 268 | my $tax_file = 'sativa_in.tax'; |
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| 269 | my $seq_file = 'sativa_in.phy'; |
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| 270 | my $res_file = 'result.mis'; |
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| 271 | |
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| 272 | my $marked_only = 0; |
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| 273 | my $mark_misplaced = 0; |
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| 274 | my $rank_test = 0; |
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| 275 | |
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| 276 | while (substr($ARGV[0],0,2) eq '--') { |
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| 277 | my $arg = shift @ARGV; |
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| 278 | if ($arg eq '--marked-only') { $marked_only = 1; } |
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| 279 | elsif ($arg eq '--mark-misplaced') { $mark_misplaced = 1; } |
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| 280 | elsif ($arg eq '--rank-test') { $rank_test = 1; } |
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| 281 | else { die_usage("Unknown switch '$arg'"); } |
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| 282 | $args--; |
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| 283 | } |
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| 284 | |
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| 285 | my $tax_field = shift @ARGV; |
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| 286 | my $species_field = shift @ARGV; |
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| 287 | my $dup_rank_names = shift @ARGV; |
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| 288 | my $wrong_rank_count = shift @ARGV; |
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| 289 | |
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| 290 | my @marklist = {}; |
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| 291 | |
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| 292 | $start_time = time; |
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| 293 | |
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| 294 | exportTaxonomy($seq_file,$tax_file,$tax_field,$species_field,$marked_only,@marklist); |
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| 295 | |
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| 296 | runPythonPipeline($seq_file,$tax_file,$dup_rank_names,$wrong_rank_count,$rank_test); |
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| 297 | |
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| 298 | importResults($res_file,$marked_only,$mark_misplaced); |
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| 299 | |
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| 300 | my $total_time = time - $start_time; |
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| 301 | print "Elapsed time: $total_time s (trainer: $trainer_time s, find_mislabels: $mislabels_time s)\n\n"; |
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| 302 | |
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| 303 | print "Done!\n\n"; |
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| 304 | } |
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| 305 | |
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| 306 | main(); |
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| 307 | |
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