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#!/usr/bin/env perl
#
# Copyright 2011, Julian Catchen <jcatchen@uoregon.edu>
#
# This file is part of Stacks.
#
# Stacks is free software: you can redistribute it and/or modify
# it under the terms of the GNU General Public License as published by
# the Free Software Foundation, either version 3 of the License, or
# (at your option) any later version.
#
# Stacks is distributed in the hope that it will be useful,
# but WITHOUT ANY WARRANTY; without even the implied warranty of
# MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE. See the
# GNU General Public License for more details.
#
# You should have received a copy of the GNU General Public License
# along with Stacks. If not, see <http://www.gnu.org/licenses/>.
#
use strict;
use DBI;
use constant stacks_version => "_VERSION_";
my $debug = 0;
my $mysql_config = "_PKGDATADIR_" . "sql/mysql.cnf";
my $db = "";
my $batch_id = 0;
my $in_path = "";
my $seq_path = "";
my $seq_type = "";
parse_command_line();
my (%sth);
prepare_sql_handles(\%sth);
if ($seq_type eq "est") {
load_ests(\%sth);
} elsif ($seq_type eq "pe_radtag") {
load_pe_radtags(\%sth);
} else {
print STDERR "Unknown sequence type specified '$seq_type'\n";
}
sub load_pe_radtags {
my ($sth) = @_;
#
# Open the file containing sequences and load them into the database.
#
my ($fh, $line, $id, $buf, $catalog_id, $seq_id, $count);
open($fh, "<$seq_path")
or die("Unable to open FASTA input file: '$seq_path', $!\n");
$count = 0;
while ($line = <$fh>) {
chomp $line;
if (substr($line, 0, 1) eq ">") {
if (length($buf) > 0) {
($catalog_id, $seq_id) = split(/\|/, $id);
$sth->{'load'}->execute($batch_id, $catalog_id, $seq_type, $seq_id, $buf)
or die("Unable to insert results into $db.\n");
$count++;
$buf = "";
}
$id = substr($line, 1);
} else {
$buf .= $line;
}
}
if (length($buf) > 0 && length($id) > 0) {
($catalog_id, $seq_id) = split(/\|/, $id);
$sth->{'load'}->execute($batch_id, $catalog_id, $seq_type, $seq_id, $buf)
or die("Unable to insert results into $db.\n");
$count++;
}
print STDERR "Loaded $count RAD-Tag paired-end contigs into the database.\n";
close($fh);
close_sql_handles(\%sth);
}
sub load_ests {
my ($sth) = @_;
#
# Load a TSV file of BLAST hits linking sequences to markers in the catalog.
#
my %hits;
load_blast_hits(\%hits);
my ($fh, $line, $id, $buf, $catalog_id, $count);
#
# Open the file containing sequences. For each sequence that has a BLAST
# hit to a marker in the catalog, load the sequence into the database.
#
open($fh, "<$seq_path")
or die("Unable to open FASTA input file: '$seq_path', $!\n");
$count = 0;
while ($line = <$fh>) {
chomp $line;
if (substr($line, 0, 1) eq ">") {
if (length($buf) > 0) {
if (defined($hits{$id})) {
foreach $catalog_id (@{$hits{$id}}) {
$sth->{'load'}->execute($batch_id, $catalog_id, $seq_type, $id, $buf)
or die("Unable to insert results into $db.\n");
$count++;
}
}
$buf = "";
}
$id = substr($line, 1);
} else {
$buf .= $line;
}
}
if (length($buf) > 0 && length($id) > 0) {
if (defined($hits{$id})) {
foreach $catalog_id (@{$hits{$id}}) {
$sth->{'load'}->execute($batch_id, $catalog_id, $seq_type, $id, $buf)
or die("Unable to insert results into $db.\n");
$count++;
}
}
}
print STDERR "Loaded $count EST sequences into the database.\n";
close($fh);
close_sql_handles(\%sth);
}
sub load_blast_hits {
my ($hits) = @_;
my (@parts, $line);
open(HITS, "<" . $in_path) or die("Unable to open BLAST hits file '$in_path' $!\n");
#
# First line is a header.
#
$line = <HITS>;
while ($line = <HITS>) {
chomp $line;
@parts = split(/\t/, $line);
if (!defined($hits->{$parts[7]})) {
$hits->{$parts[7]} = [];
}
if (!grep(/^$parts[2]$/, @{$hits->{$parts[7]}})) {
push(@{$hits->{$parts[7]}}, $parts[2]);
}
}
print STDERR "Loaded ", scalar(keys %{$hits}), " distinct BLAST hits.\n";
close(HITS);
}
sub prepare_sql_handles {
my ($sth) = @_;
my $cnf = (defined($ENV{"HOME"}) && -e $ENV{"HOME"} . "/.my.cnf") ? $ENV{"HOME"} . "/.my.cnf" : $mysql_config;
$sth->{'dbh'} = DBI->connect("DBI:mysql:$db:mysql_read_default_file=$cnf")
or die("Unable to connect to the $db MySQL Database!\n" . $DBI::errstr);
my $query;
$query =
"INSERT INTO sequence SET batch_id=?, catalog_id=?, type=?, seq_id=?, seq=?";
$sth->{'load'} = $sth->{'dbh'}->prepare($query) or die($sth->{'dbh'}->errstr());
}
sub close_sql_handles {
my ($sth) = @_;
my $key;
foreach $key (keys %{$sth}) {
next if ($key =~ /dbh/);
$sth->{$key}->finish();
}
foreach $key (keys %{$sth}) {
next if ($key !~ /dbh/);
$sth->{$key}->disconnect();
}
}
sub parse_command_line {
while (@ARGV) {
$_ = shift @ARGV;
if ($_ =~ /^-p$/) { $in_path = shift @ARGV; }
elsif ($_ =~ /^-f$/) { $seq_path = shift @ARGV; }
elsif ($_ =~ /^-b$/) { $batch_id = shift @ARGV; }
elsif ($_ =~ /^-D$/) { $db = shift @ARGV; }
elsif ($_ =~ /^-t$/) { $seq_type = shift @ARGV; }
elsif ($_ =~ /^-d$/) { $debug++; }
elsif ($_ =~ /^-h$/) { usage(); }
else {
print STDERR "Unknown command line option: '$_'\n";
usage();
}
}
if (length($seq_type) == 0) {
print STDERR "You must specify the sequence type.\n";
usage();
}
if ($batch_id == 0) {
print STDERR "You must specify the batch ID.\n";
usage();
}
}
sub version {
print STDERR "load_sequences.pl ", stacks_version, "\n";
}
sub usage {
version();
print STDERR <<EOQ;
load_sequences.pl -b id -D db -f path -t type [-p blast_path] [-d] [-h]
b: batch ID.
D: database to examine.
f: path to FASTA file containing sequences to load.
t: sequence type to load: either 'pe_radtag' or 'est'.
p: path to tab-separated file containing BLAST hits.
h: display this help message.
d: turn on debug output.
EOQ
exit(0);
}
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