Usage of Perl Needleman/Wunsch Module
2
0
Entering edit mode
10.4 years ago
biolab ★ 1.4k

Dear all,

I installed Needleman-Wunsch module, however have some problems to handle it. Could anyone help to write a simple example to illustrate how to use it? My second question is: two sequences could align in different forms, my work actually only needs the best alignment, however I'd like to know how to output the other forms. Where is the parameter to do this? MY third question is: From literature I know the score normally is set as follows, match:1, mismatch:-1, gap:-2, then what's gap extention and gap open? WHat's your favorite score scheme? Note I am aligning nucleotide sequences.

I will much appreciate your help if you answer any of my three questions. THANK YOU!!

SYNOPSIS

use Algorithm::NeedlemanWunsch;
sub score_sub {
    if (!@_) {
        return -2; # gap penalty
    }

    return ($_[0] eq $_[1]) ? 1 : -1;
}
my $matcher = Algorithm::NeedlemanWunsch->new(\&score_sub);
my $score = $matcher->align(
    \@a,
    \@b,
    { align => \&on_align,
        shift_a => \&on_shift_a,
        shift_b => \&on_shift_b,
        select_align => \&on_select_align
    });

DESCRIPTION

Sequence alignment is a way to find commonalities in two (or more) similar sequences or strings of some items or characters. Standard motivating example is the comparison of DNA sequences and their functional and evolutionary similarities and differences, but the problem has much wider applicability - for example finding the longest common subsequence (that is, "diff") is a special case of sequence alignment. ... ...

METHODS

Standard algorithm

new(\&score_sub [, $gap_penalty ])

The constructor. Takes one mandatory argument, which is a coderef to a sub implementing the similarity matrix, plus an optional gap penalty argument. If the gap penalty isn't specified as a constructor argument, the "Algorithm::NeedlemanWunsch" object gets it by calling the scoring sub without arguments; apart from that case, the sub is called with 2 arguments, which are items from the first and second sequence, respectively, passed to "Algorithm::NeedlemanWunsch::align". Note that the sub must be pure, i.e. always return the same value when called with the same arguments.

align(\@a, \@b [, \%callbacks ])

The core of the algorithm. Creates a bottom-up dynamic programming matrix, fills it with alignment scores and then traces back to find an optimal alignment, informing the application about its items by invoking the callbacks passed to the method.

The first 2 arguments of "align" are array references to the aligned sequences, the third a hash reference with user-supplied callbacks. The callbacks are identified by the hash keys, which are as follows:

  • align: Aligns two sequence items. The callback is called with 2 arguments, which are the positions of the paired items in "\@a" and "\@b", respectively.
  • shift_a: Aligns an item of the first sequence with a gap in the second sequence. The callback is called with 1 argument, which is the position of the item in "\@a".
  • shift_b: Aligns a gap in the first sequence with an item of the second sequence. The callback is called with 1 argument, which is the position of the item in "\@b".
  • select_align: Called when there's more than one way to construct the optimal alignment, with 1 argument which is a hashref enumerating the possibilities. The hash may contain the following keys:

    • align: If this key exists, the optimal alignment may align two sequence items. The key's value is an arrayref with the positions of the paired items in "\@a" and "\@b", respectively.
    • shift_a: If this key exists, the optimal alignment may align an item of the first sequence with a gap in the second sequence. The key's value is the position of the item in "\@a".
    • shift_b: If this key exists, the optimal alignment may align a gap in the first sequence with an item of the second sequence. The key's value is the position of the item in "\@b".

    All keys are optional, but the hash will always have at least one.The callback must select one of the possibilities by returning one of the keys.

All callbacks are optional. When there is just one way to make the optimal alignment, the "Algorithm::NeedlemanWunsch" object prefers calling the specific callbacks, but will call "select_align" if it's defined and the specific callback isn't.

Note that "select_align" is called instead of the specific callbacks, not in addition to them - users defining both "select_align" and other callbacks should probably call the specific callback explicitly from their "select_align", once it decides which one to prefer.

Also note that the passed positions move backwards, from the sequence ends to zero - if you're building the alignment in your callbacks, add items to the front. ... ...

alignment global needleman-wunsh PERL module • 7.1k views
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2
Entering edit mode

I think you are asking a lot here, I would recommend limiting the scope of your post to one or two questions and try to provide some relevant information about the errors (not just copying all the documentation). What exactly are you trying to do and what problems did you have with the module? Also, what do you want as output, the alignment or the scores, or something else maybe?

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0
Entering edit mode

Hi SES, thanks for your advices. I clicked ADD COMMENT or ADD REPLY, but did not work (I tried IE and Firefox), I have to use this answer box. Regarding to my original question, I mostly concern how to use this module. Could anyone give me an example of using it? For instance, $seq1 = 'ATGCATGCATGC' $seq2 = 'ATGGATCGAGC'. I am new in perl, especially handle modules. Thank you very much!

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2
Entering edit mode
10.4 years ago
Michael 55k

Here is an example that works and is more instructive than the Synopsis section to explain how to use the package:


#!/usr/bin/env perl

use strict;
use warnings;
use Algorithm::NeedlemanWunsch;
### scoring function:
sub score_sub {
  if (!@_) {
    return -2;            # gap penalty
  }
  ## mismatch scores -1, match +1
  return ($_[0] eq $_[1]) ? 1 : -1;
} 

# sequences come as strings normally
my $a = "ATCTATC";
my $b = "GTCTTTTTTTTATC";
my @a = split //, $a; # make an array
my @b = split //, $b;

## callbacks that print something useful
## prints an 'alignment string' in the order of the  
## recursion of the dynamic programming algorithm 
## print "-" only on match
sub on_align  { print "align", " " , $a[$_[0]], ($a[$_[0]] eq $b[$_[1]] ) ? "-" : " ", $b[$_[1]], "\n" }; 
sub on_shift_a {  print "gap  ", "" , $a[$_[0]], "\n" };
sub on_shift_b { print "gap  ", "   " , $b[$_[0]], "\n"};
### Dumb select, need to return one of the keys for alternative 
### alignments with equal score. Here, we always take the first option, but don't print it.

sub on_select_align { print "(select_align)\n"; return (keys (%{$_[0]})) [0]};
## one gets the same behaviour with not assigning on_select_align, I am too lazy to implement this callback correctly ...

my $matcher = Algorithm::NeedlemanWunsch->new(\&score_sub);
my $score = $matcher->align(
                \@a,
                \@b,
                {   align     => \&on_align,
                shift_a => \&on_shift_a,
                shift_b => \&on_shift_b,
 #               select_align => \&on_select_align
                });

print "score: $score\n";

Output:

align C-C
align T-T
align A-A
align T-T
gap     T
gap     T
gap     T
gap     T
gap     T
gap     T
gap     T
align C-C
align T-T
align A G
score: -9
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0
Entering edit mode

Hi Mike, Thank you for your detailed perl script that is much helpful for my work. THANKS!

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0
Entering edit mode

Dear Mike, I am trying to modify your perl script to change output format (I would like to output two aligned sequences on a single line). Unfortunately I had troubles. Could you please check this script? THANKS!

#!/usr/bin/env perl

use strict;
use warnings;
use Algorithm::NeedlemanWunsch;
### scoring function:
sub score_sub {
  if (!@_) {
    return -2;            # gap penalty
  }
  ## mismatch scores -1, match +1
  return ($_[0] eq $_[1]) ? 1 : -1;
}


my $a = "ATCTATC";
my $b = "GTCTTTTTTTTATC";
my @a = split //, $a; # make an array
my @b = split //, $b;

my ($a_align, $b_align);#


## callbacks that print something useful
## prints an 'alignment string' in the order of the  
## recursion of the dynamic programming algorithm
## print "-" only on match
sub on_align  {  
$a_align .= $a[$_[0]];
$b_align .= $b[$_[1]];
};

sub on_shift_a { $a_align .= $a[$_[0]];};
sub on_shift_b { $b_align .= $b[$_[0]];};

### Dumb select, need to return one of the keys for alternative
### alignments with equal score. Here, we always take the first option, but don't print it.

sub on_select_align { print "(select_align)\n"; return (keys (%{$_[0]})) [0]};
## one gets the same behaviour with not assigning on_select_align, I am too lazy to implement this callback correctly ...

my $matcher = Algorithm::NeedlemanWunsch->new(\&score_sub);
my $score = $matcher->align(
                \@a,
                \@b,
                {   align     => \&on_align,
                shift_a => \&on_shift_a,
                shift_b => \&on_shift_b,
         #select_align => \&on_select_align
                });

print "$a_align\t$b_align\t";
print "$score\n";

My ideal output is :

CTAT-------CTA   CTATTTTTTTTCTG   -9
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1
Entering edit mode

Hi Biolab,

you need to change the on_shift_a and on_shift_b subroutines:

sub on_shift_a { $a_align .= $a[$_[0]]; $b_align .= '-'};
sub on_shift_b { $b_align .= $b[$_[0]]; $a_align .= '-'};
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0
Entering edit mode

I've followed this thread and it's really helpful, thanks. Can someone indicate to me how to use the module Extensions gap_open_penalty and gap_extend_penalty in order to implement affine gap penalties? (The stuff described here)

Thanks for any help,
Nick Goldman

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0
Entering edit mode
$matcher->gap_open_penalty($gappen); $matcher->gap_extend_penalty($gapextpen);

Before calling matcher->align(); This works very well but then I tried to implement it over a loop, with $a and $b different sequences each time, it says: Use of uninitialized value $a_align in concatenation (.) or string. Same for $b_align. The callback with print shows the algorithm is working. Anyone has any idea as to how to re-initialize $a_align and $b_align?

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0
Entering edit mode

I found this on a russian forum:

my (@align1, @align2);
my $result = $matcher->align($arr1, $arr2,
  {
   align   => sub {unshift @align1, $arr1->[shift]; unshift @align2, $arr2->[shift]},
   shift_a => sub {unshift @align1, $arr1->[shift]; unshift @align2,            '.'},
   shift_b => sub {unshift @align1,            '.'; unshift @align2, $arr1->[shift]},
  });

$arr1 and $arr2 are references to the array of sequences that you would get as

my $arr1 = [split //, $a];

Worked for me.

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0
Entering edit mode
6.5 years ago

$matcher->gap_open_penalty(-2); $matcher->gap_extend_penalty(-0.1); Before calling matcher->align(); This works very well but then I tried to implement it over a loop, with $a and $b different sequences each time, it says: Use of uninitialized value $a_align in concatenation (.) or string. Same for $b_align. The callback with print shows the algorithm is working. Anyone has any idea as to how to re-initialize $a_align and $b_align?

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0
Entering edit mode

I found this on a russian forum:

my (@align1, @align2);
my $result = $matcher->align($arr1, $arr2,
  {
   align   => sub {unshift @align1, $arr1->[shift]; unshift @align2, $arr2->[shift]},
   shift_a => sub {unshift @align1, $arr1->[shift]; unshift @align2,            '.'},
   shift_b => sub {unshift @align1,            '.'; unshift @align2, $arr1->[shift]},
  });

$arr1 and $arr2 are references to the array of sequences that you would get as

my $arr1 = [split //, $a];

Worked for me.

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