NAME
Bio::DB::GFF -- Storage and retrieval of sequence annotation data
SYNOPSIS
use Bio::DB::GFF;
# Open the sequence database
my $db = Bio::DB::GFF->new( -adaptor => 'dbi::mysqlopt',
-dsn => 'dbi:mysql:elegans',
-fasta => '/usr/local/fasta_files'
);
# fetch a 1 megabase segment of sequence starting at landmark "ZK909"
my $segment = $db->segment('ZK909', 1 => 1000000);
# pull out all transcript features
my @transcripts = $segment->features('transcript');
# for each transcript, total the length of the introns
my %totals;
for my $t (@transcripts) {
my @introns = $t->Intron;
$totals{$t->name} += $_->length foreach @introns;
}
# Sort the exons of the first transcript by position
my @exons = sort {$a->start <=> $b->start} $transcripts[0]->Exon;
# Get a region 1000 bp upstream of first exon
my $upstream = $exons[0]->segment(-1000,0);
# get its DNA
my $dna = $upstream->dna;
# and get all curated polymorphisms inside it
@polymorphisms = $upstream->contained_features('polymorphism:curated');
# get all feature types in the database
my @types = $db->types;
# count all feature types in the segment
my %type_counts = $segment->types(-enumerate=>1);
# get an iterator on all curated features of type 'exon' or 'intron'
my $iterator = $db->get_seq_stream(-type => ['exon:curated','intron:curated']);
while (my $s = $iterator->next_seq) {
print $s,"\n";
}
# find all transcripts annotated as having function 'kinase'
my $iterator = $db->get_seq_stream(-type=>'transcript',
-attributes=>{Function=>'kinase'});
while (my $s = $iterator->next_seq) {
print $s,"\n";
}
DESCRIPTION
Bio::DB::GFF provides fast indexed access to a sequence annotation database. It supports multiple database types (ACeDB, relational), and multiple schemas through a system of adaptors and aggregators.
The following operations are supported by this module:
- retrieving a segment of sequence based on the ID of a landmark
- retrieving the DNA from that segment
- finding all annotations that overlap with the segment
- finding all annotations that are completely contained within the
segment
- retrieving all annotations of a particular type, either within a
segment, or globally
- conversion from absolute to relative coordinates and back again,
using any arbitrary landmark for the relative coordinates
- using a sequence segment to create new segments based on relative
offsets
The data model used by Bio::DB::GFF is compatible with the GFF flat file format (http://www.sanger.ac.uk/software/GFF). The module can load a set of GFF files into the database, and serves objects that have methods corresponding to GFF fields.
The objects returned by Bio::DB::GFF are compatible with the SeqFeatureI interface, allowing their use by the Bio::Graphics and Bio::DAS modules.
Auxiliary Scripts
The bioperl distribution includes several scripts that make it easier to work with Bio::DB::GFF databases. They are located in the scripts directory under a subdirectory named Bio::DB::GFF:
- load_gff.pl
-
This script will load a Bio::DB::GFF database from a flat GFF file of sequence annotations. Only the relational database version of Bio::DB::GFF is supported. It can be used to create the database from scratch, as well as to incrementally load new data.
This script takes a --fasta argument to load raw DNA into the database as well. However, GFF databases do not require access to the raw DNA for most of their functionality.
load_gff.pl also has a --upgrade option, which will perform a non-destructive upgrade of older schemas to newer ones.
- bulk_load_gff.pl
-
This script will populate a Bio::DB::GFF database from a flat GFF file of sequence annotations. Only the MySQL database version of Bio::DB::GFF is supported. It uses the "LOAD DATA INFILE" query in order to accelerate loading considerably; however, it can only be used for the initial load, and not for updates.
This script takes a --fasta argument to load raw DNA into the database as well. However, GFF databases do not require access to the raw DNA for most of their functionality.
- gadfly_to_gff.pl
-
This script will convert the GFF-like format used by the Berkeley Drosophila Sequencing project into a format suitable for use with this module.
- sgd_to_gff.pl
-
This script will convert the tab-delimited feature files used by the Saccharomyces Genome Database into a format suitable for use with this module.
GFF Fundamentals
The GFF format is a flat tab-delimited file, each line of which corresponds to an annotation, or feature. Each line has nine columns and looks like this:
Chr1 curated CDS 365647 365963 . + 1 Transcript "R119.7"
The 9 columns are as follows:
- 1. reference sequence
-
This is the ID of the sequence that is used to establish the coordinate system of the annotation. In the example above, the reference sequence is "Chr1".
- 2. source
-
The source of the annotation. This field describes how the annotation was derived. In the example above, the source is "curated" to indicate that the feature is the result of human curation. The names and versions of software programs are often used for the source field, as in "tRNAScan-SE/1.2".
- 3. method
-
The annotation method. This field describes the type of the annotation, such as "CDS". Together the method and source describe the annotation type.
- 4. start position
-
The start of the annotation relative to the reference sequence.
- 5. stop position
-
The stop of the annotation relative to the reference sequence. Start is always less than or equal to stop.
- 6. score
-
For annotations that are associated with a numeric score (for example, a sequence similarity), this field describes the score. The score units are completely unspecified, but for sequence similarities, it is typically percent identity. Annotations that don't have a score can use "."
- 7. strand
-
For those annotations which are strand-specific, this field is the strand on which the annotation resides. It is "+" for the forward strand, "-" for the reverse strand, or "." for annotations that are not stranded.
- 8. phase
-
For annotations that are linked to proteins, this field describes the phase of the annotation on the codons. It is a number from 0 to 2, or "." for features that have no phase.
- 9. group
-
GFF provides a simple way of generating annotation hierarchies ("is composed of" relationships) by providing a group field. The group field contains the class and ID of an annotation which is the logical parent of the current one. In the example given above, the group is the Transcript named "R119.7".
The group field is also used to store information about the target of sequence similarity hits, and miscellaneous notes. See the next section for a description of how to describe similarity targets.
The sequences used to establish the coordinate system for annotations can correspond to sequenced clones, clone fragments, contigs or super-contigs. Thus, this module can be used throughout the lifecycle of a sequencing project.
In addition to a group ID, the GFF format allows annotations to have a group class. For example, in the ACeDB representation, RNA interference experiments have a class of "RNAi" and an ID that is unique among the RNAi experiments. Since not all databases support this notion, the class is optional in all calls to this module, and defaults to "Sequence" when not provided.
Double-quotes are sometimes used in GFF files around components of the group field. Strictly, this is only necessary if the group name or class contains whitespace.
Making GFF files work with this module
Some annotations do not need to be individually named. For example, it is probably not useful to assign a unique name to each ALU repeat in a vertebrate genome. Others, such as predicted genes, correspond to named biological objects; you probably want to be able to fetch the positions of these objects by referring to them by name.
To accomodate named annotations, the GFF format places the object class and name in the group field. The name identifies the object, and the class prevents similarly-named objects, for example clones and sequences, from collding.
A named object is shown in the following excerpt from a GFF file:
Chr1 curated transcript 939627 942410 . + . Transcript Y95B8A.2
This object is a predicted transcript named Y95BA.2. In this case, the group field is used to identify the class and name of the object, even though no other annotation belongs to that group.
It now becomes possible to retrieve the region of the genome covered by transcript Y95B8A.2 using the segment() method:
$segment = $db->segment(-class=>'Transcript',-name=>'Y95B8A.2');
It is not necessary for the annotation's method to correspond to the object class, although this is commonly the case.
As explained above, each annotation in a GFF file refers to a reference sequence. It is important that each reference sequence also be identified by a line in the GFF file. This allows the Bio::DB::GFF module to determine the length and class of the reference sequence, and makes it possible to do relative arithmetic.
For example, if "Chr1" is used as a reference sequence, then it should have an entry in the GFF file similar to this one:
Chr1 assembly chromosome 1 14972282 . + . Sequence Chr1
This indicates that the reference sequence named "Chr1" has length 14972282 bp, method "chromosome" and source "assembly". In addition, as indicated by the group field, Chr1 has class "Sequence" and name "Chr".
The object class "Sequence" is used by default when the class is not specified in the segment() call. This allows you to use a shortcut form of the segment() method:
$segment = $db->segment('Chr1'); # whole chromosome
$segment = $db->segment('Chr1',1=>1000); # first 1000 bp
For your convenience, if, during loading a GFF file, Bio::DB::GFF encounters a line like the following:
##sequence-region Chr1 1 14972282
It will automatically generate the following entry:
Chr1 reference Component 1 14972282 . + . Sequence Chr1
This is sufficient to use Chr1 as a reference point. The ##sequence-region line is frequently found in the GFF files distributed by annotation groups.
Sequence alignments
There are two cases in which an annotation indicates the relationship between two sequences. The first case is a similarity hit, where the annotation indicates an alignment. The second case is a map assembly, in which the annotation indicates that a portion of a larger sequence is built up from one or more smaller ones.
Both cases are indicated by using the Target tag in the group field. For example, a typical similarity hit will look like this:
Chr1 BLASTX similarity 76953 77108 132 + 0 Target Protein:SW:ABL_DROME 493 544
The group field contains the Target tag, followed by an identifier for the biological object referred to. The GFF format uses the notation Class:Name for the biological object, and even though this is stylistically inconsistent, that's the way it's done. The object identifier is followed by two integers indicating the start and stop of the alignment on the target sequence.
Unlike the main start and stop columns, it is possible for the target start to be greater than the target end. The previous example indicates that the the section of Chr1 from 76,953 to 77,108 aligns to the protein SW:ABL_DROME starting at position 493 and extending to position 544.
A similar notation is used for sequence assembly information as shown in this example:
Chr1 assembly Link 10922906 11177731 . . . Target Sequence:LINK_H06O01 1 254826
LINK_H06O01 assembly Cosmid 32386 64122 . . . Target Sequence:F49B2 6 31742
This indicates that the region between bases 10922906 and 11177731 of Chr1 are composed of LINK_H06O01 from bp 1 to bp 254826. The region of LINK_H0601 between 32386 and 64122 is, in turn, composed of the bases 5 to 31742 of cosmid F49B2.
Attributes
While not intended to serve as a general-purpose sequence database (see bioperl-db for that), GFF allows you to tag features with arbitrary attributes. Attributes appear in the Group field following the initial class/name pair. For example:
Chr1 cur trans 939 942 . + . Transcript Y95B8A.2 ; Gene sma-3 ; Alias sma3
This line tags the feature named Transcript Y95B8A.2 as being "Gene" named sma-3 and having the Alias "sma3". Features having these attributes can be looked up using the fetch_feature_by_attribute() method.
Two attributes have special meaning: "Note" is for backward compatibility and is used for unstructured text remarks. "Alias" is considered as a synonym for the feature name and will be consulted when looking up a feature by its name.
Adaptors and Aggregators
This module uses a system of adaptors and aggregators in order to make it adaptable to use with a variety of databases.
- Adaptors
-
The core of the module handles the user API, annotation coordinate arithmetic, and other common issues. The details of fetching information from databases is handled by an adaptor, which is specified during Bio::DB::GFF construction. The adaptor encapsulates database-specific information such as the schema, user authentication and access methods.
Currently there are two adaptors: 'dbi::mysql' and 'dbi::mysqlopt'. The former is an interface to a simple Mysql schema. The latter is an optimized version of dbi::mysql which uses a binning scheme to accelerate range queries and the Bio::DB::Fasta module for rapid retrieval of sequences. Note the double-colon between the words.
- Aggregators
-
The GFF format uses a "group" field to indicate aggregation properties of individual features. For example, a set of exons and introns may share a common transcript group, and multiple transcripts may share the same gene group.
Aggregators are small modules that use the group information to rebuild the hierarchy. When a Bio::DB::GFF object is created, you indicate that it use a set of one or more aggregators. Each aggregator provides a new composite annotation type. Before the database query is generated each aggregator is called to "disaggregate" its annotation type into list of component types contained in the database. After the query is generated, each aggregator is called again in order to build composite annotations from the returned components.
For example, during disaggregation, the standard "transcript" aggregator generates a list of component feature types including "intron", "exon", "CDS", and "3'UTR". Later, it aggregates these features into a set of annotations of type "transcript".
During aggregation, the list of aggregators is called in reverse order. This allows aggregators to collaborate to create multi-level structures: the transcript aggregator assembles transcripts from introns and exons; the gene aggregator then assembles genes from sets of transcripts.
Three default aggregators are provided:
transcript assembles transcripts from features of type exon, CDS, 5'UTR, 3'UTR, TSS, and PolyA clone assembles clones from Clone_left_end, Clone_right_end and Sequence features. alignment assembles gapped alignments from features of type "similarity".
In addition, this module provides the optional "wormbase_gene" aggregator, which accomodates the WormBase representation of genes. This aggregator aggregates features of method "exon", "CDS", "5'UTR", "3'UTR", "polyA" and "TSS" into a single object. It also expects to find a single feature of type "Sequence" that spans the entire gene.
The existing aggregators are easily customized.
Note that aggregation will not occur unless you specifically request the aggregation type. For example, this call:
@features = $segment->features('alignment');
will generate an array of aggregated alignment features. However, this call:
@features = $segment->features();
will return a list of unaggregated similarity segments.
For more informnation, see the manual pages for Bio::DB::GFF::Aggregator::transcript, Bio::DB::GFF::Aggregator::clone, etc.
API
The following is the API for Bio::DB::GFF.
Querying GFF Databases
new
Title : new
Usage : my $db = new Bio::DB::GFF(@args);
Function: create a new Bio::DB::GFF object
Returns : new Bio::DB::GFF object
Args : lists of adaptors and aggregators
Status : Public
These are the arguments:
-adaptor Name of the adaptor module to use. If none
provided, defaults to "dbi:mysqlopt".
-aggregator Array reference to a list of aggregators
to apply to the database. If none provided,
defaults to ['transcript','clone','alignment'].
<other> Any other named argument pairs are passed to
the adaptor for processing.
The adaptor argument must correspond to a module contained within the Bio::DB::GFF::Adaptor namespace. For example, the Bio::DB::GFF::Adaptor::dbi::mysql adaptor is loaded by specifying 'dbi:mysql'. By Perl convention, the adaptors names are lower case because they are loaded at run time.
The aggregator array may contain a list of aggregator names, or a list of initialized aggregator objects. For example, if you wish to change the components aggregated by the transcript aggregator, you could pass it to the GFF constructor this way:
my $transcript =
Bio::DB::Aggregator::transcript->new(-sub_parts=>[qw(exon intron utr
polyA spliced_leader)]);
my $db = Bio::DB::GFF->new(-aggregator=>[$transcript,'clone','alignment],
-adaptor => 'dbi:mysql',
-dsn => 'dbi:mysql:elegans42');
The commonly used 'dbi:mysql' adaptor recognizes the following adaptor-specific arguments:
Argument Description
-------- -----------
-dsn the DBI data source, e.g. 'dbi:mysql:ens0040'
If a partial name is given, such as "ens0040", the
"dbi:mysql:" prefix will be added automatically.
-user username for authentication
-pass the password for authentication
The commonly used 'dbi:mysqlopt' adaptor also recogizes the following arguments.
Argument Description
-------- -----------
-fasta path to a directory containing FASTA files for the DNA
contained in this database (e.g. "/usr/local/share/fasta")
-acedb an acedb URL to use when converting features into ACEDB
objects (e.g. sace://localhost:2005)
types
Title : types
Usage : $db->types(@args)
Function: return list of feature types in range or database
Returns : a list of Bio::DB::GFF::Typename objects
Args : see below
Status : public
This routine returns a list of feature types known to the database. The list can be database-wide or restricted to a region. It is also possible to find out how many times each feature occurs.
For range queries, it is usually more convenient to create a Bio::DB::GFF::Segment object, and then invoke it's types() method.
Arguments are as follows:
-ref ID of reference sequence
-class class of reference sequence
-start start of segment
-stop stop of segment
-enumerate if true, count the features
The returned value will be a list of Bio::DB::GFF::Typename objects, which if evaluated in a string context will return the feature type in "method:source" format. This object class also has method() and source() methods for retrieving the like-named fields.
If -enumerate is true, then the function returns a hash (not a hash reference) in which the keys are type names in "method:source" format and the values are the number of times each feature appears in the database or segment.
The argument -end is a synonum for -stop, and -count is a synonym for -enumerate.
classes
Title : classes
Usage : $db->classes
Function: return list of landmark classes in database
Returns : a list of classes
Args : none
Status : public
This routine returns the list of reference classes known to the database, or empty if classes are not used by the database. Classes are distinct from types, being essentially qualifiers on the reference namespaces.
segment
Title : segment
Usage : $db->segment(@args);
Function: create a segment object
Returns : segment object(s)
Args : numerous, see below
Status : public
This method generates a segment object, which is a Perl object subclassed from Bio::DB::GFF::Segment. The segment can be used to find overlapping features and the raw DNA.
When making the segment() call, you specify the ID of a sequence landmark (e.g. an accession number, a clone or contig), and a positional range relative to the landmark. If no range is specified, then the entire extent of the landmark is used to generate the segment.
You may also provide the ID of a "reference" sequence, which will set the coordinate system and orientation used for all features contained within the segment. The reference sequence can be changed later. If no reference sequence is provided, then the coordinate system is based on the landmark.
Arguments:
-name ID of the landmark sequence.
-class Database object class for the landmark sequence.
"Sequence" assumed if not specified. This is
irrelevant for databases which do not recognize
object classes.
-start Start of the segment relative to landmark. Positions
follow standard 1-based sequence rules. If not specified,
defaults to the beginning of the landmark.
-end Stop of the segment relative to the landmark. If not specified,
defaults to the end of the landmark.
-stop Same as -end.
-offset For those who prefer 0-based indexing, the offset specifies the
position of the new segment relative to the start of the landmark.
-length For those who prefer 0-based indexing, the length specifies the
length of the new segment.
-refseq Specifies the ID of the reference landmark used to establish the
coordinate system for the newly-created segment.
-refclass Specifies the class of the reference landmark, for those databases
that distinguish different object classes. Defaults to "Sequence".
-absolute
Return features in absolute coordinates rather than relative to the
parent segment.
-nocheck Don't check the database for the coordinates and length of this
feature. Construct a segment using the indicated name as the
reference, a start coordinate of 1, an undefined end coordinate,
and a strand of +1.
-force Same as -nocheck.
-seq,-sequence,-sourceseq Aliases for -name.
-begin,-end Aliases for -start and -stop
-off,-len Aliases for -offset and -length
-seqclass Alias for -class
Here's an example to explain how this works:
my $db = Bio::DB::GFF->new(-dsn => 'dbi:mysql:human',-adaptor=>'dbi:mysql');
If successful, $db will now hold the database accessor object. We now try to fetch the fragment of sequence whose ID is A0000182 and class is "Accession."
my $segment = $db->segment(-name=>'A0000182',-class=>'Accession');
If successful, $segment now holds the entire segment corresponding to this accession number. By default, the sequence is used as its own reference sequence, so its first base will be 1 and its last base will be the length of the accession.
Assuming that this sequence belongs to a longer stretch of DNA, say a contig, we can fetch this information like so:
my $sourceseq = $segment->sourceseq;
and find the start and stop on the source like this:
my $start = $segment->abs_start;
my $stop = $segment->abs_stop;
If we had another segment, say $s2, which is on the same contiguous piece of DNA, we can pass that to the refseq() method in order to establish it as the coordinat reference point:
$segment->refseq($s2);
Now calling start() will return the start of the segment relative to the beginning of $s2, accounting for differences in strandedness:
my $rel_start = $segment->start;
IMPORTANT NOTE: This method can be used to return the segment spanned by an arbitrary named annotation. However, if the annotation appears at multiple locations on the genome, for example an EST that maps to multiple locations, then, provided that all locations reside on the same physical segment, the method will return a segment that spans the minimum and maximum positions. If the reference sequence occupies ranges on different physical segments, then it returns them all in an array context, and raises a "multiple segment exception" exception in a scalar context.
features
Title : features
Usage : $db->features(@args)
Function: get all features, possibly filtered by type
Returns : a list of Bio::DB::GFF::Feature objects
Args : see below
Status : public
This routine will retrieve features in the database regardless of position. It can be used to return all features, or a subset based on their method and source.
Arguments are as follows:
-types List of feature types to return. Argument is an array
reference containing strings of the format "method:source"
-merge Whether to apply aggregators to the generated features.
-rare Turn on optimizations suitable for a relatively rare feature type,
where it makes more sense to filter by feature type first,
and then by position.
-attributes A hash reference containing attributes to match.
-iterator Whether to return an iterator across the features.
If -iterator is true, then the method returns a single scalar value consisting of a Bio::SeqIO object. You can call next_seq() repeatedly on this object to fetch each of the features in turn. If iterator is false or absent, then all the features are returned as a list.
Currently aggregation is disabled when iterating over a series of features.
Types are indicated using the nomenclature "method:source". Either of these fields can be omitted, in which case a wildcard is used for the missing field. Type names without the colon (e.g. "exon") are interpreted as the method name and a source wild card. Regular expressions are allowed in either field, as in: "similarity:BLAST.*".
The -attributes argument is a hashref containing one or more attributes to match against:
-attributes => { Gene => 'abc-1',
Note => 'confirmed' }
Attribute matching is simple string matching, and multiple attributes are ANDed together.
get_seq_stream
Title : get_seq_stream
Usage : my $seqio = $self->get_seq_sream(@args)
Function: Performs a query and returns an iterator over it
Returns : a Bio::SeqIO stream capable of producing sequence
Args : As in features()
Status : public
This routine takes the same arguments as features(), but returns a Bio::SeqIO::Stream-compliant object. Use it like this:
$stream = $db->get_seq_stream('exon');
while (my $exon = $stream->next_seq) {
print $exon,"\n";
}
NOTE: This is also called get_feature_stream(), since that's what it really does.
get_feature_by_name
Title : get_feature_by_name
Usage : $db->get_feature_by_name($class => $name)
Function: fetch features by their name
Returns : a list of Bio::DB::GFF::Feature objects
Args : the class and name of the desired feature
Status : public
This method can be used to fetch a named feature from the database. GFF annotations are named using the group class and name fields, so for features that belong to a group of size one, this method can be used to retrieve that group (and is equivalent to the segment() method).
This method may return zero, one, or several Bio::DB::GFF::Feature objects.
Aggregation is performed on features as usual.
NOTE: At various times, this function was called fetch_group(), fetch_feature(), fetch_feature_by_name() and segments(). These names are preserved for backward compatibility.
get_feature_by_target
Title : get_feature_by_target
Usage : $db->get_feature_by_target($class => $name)
Function: fetch features by their similarity target
Returns : a list of Bio::DB::GFF::Feature objects
Args : the class and name of the desired feature
Status : public
This method can be used to fetch a named feature from the database based on its similarity hit. In this implementation it is identical to get_feature_by_target().
get_feature_by_attribute
Title : get_feature_by_attribute
Usage : $db->get_feature_by_attribute(attribute1=>value1,attribute2=>value2)
Function: fetch segments by combinations of attribute values
Returns : a list of Bio::DB::GFF::Feature objects
Args : the class and name of the desired feature
Status : public
This method can be used to fetch a set of features from the database. Attributes are a list of name=>value pairs. They will be logically ANDED together.
get_feature_by_id
Title : get_feature_by_id
Usage : $db->get_feature_by_id($id)
Function: fetch segments by feature ID
Returns : a Bio::DB::GFF::Feature object
Args : the feature ID
Status : public
This method can be used to fetch a feature from the database using its ID. Not all GFF databases support IDs, so be careful with this.
get_feature_by_gid
Title : get_feature_by_gid
Usage : $db->get_feature_by_gid($id)
Function: fetch segments by feature ID
Returns : a Bio::DB::GFF::Feature object
Args : the feature ID
Status : public
This method can be used to fetch a feature from the database using its group ID. Not all GFF databases support IDs, so be careful with this.
The group ID is often more interesting than the feature ID, since groups can be complex objects containing subobjects.
absolute
Title : absolute
Usage : $abs = $db->absolute([$abs]);
Function: gets/sets absolute mode
Returns : current setting of absolute mode boolean
Args : new setting for absolute mode boolean
Status : public
$db->absolute(1) will turn on absolute mode for the entire database. All segments retrieved will use absolute coordinates by default, rather than relative coordinates. You can still set them to use relative coordinates by calling $segment->absolute(0).
Note that this is not the same as calling abs_segment(); it continues to allow you to look up groups that are not used directly as reference sequences.
strict_bounds_checking
Title : strict_bounds_checking
Usage : $flag = $db->strict_bounds_checking([$flag])
Function: gets/sets strict bounds checking
Returns : current setting of bounds checking flag
Args : new setting for bounds checking flag
Status : public
This flag enables extra checks for segment requests that go beyond the ends of their reference sequences. If bounds checking is enabled, then retrieved segments will be truncated to their physical length, and their truncated() methods will return true.
If the flag is off (the default), then the module will return segments that appear to extend beyond their physical boundaries. Requests for features beyond the end of the segment will, however, return empty.
get_Seq_by_id
Title : get_Seq_by_id
Usage : $seq = $db->get_Seq_by_id('ROA1_HUMAN')
Function: Gets a Bio::Seq object by its name
Returns : a Bio::Seq object
Args : the id (as a string) of a sequence
Throws : "id does not exist" exception
NOTE: Bio::DB::RandomAccessI compliant method
get_Seq_by_accession
Title : get_Seq_by_accession
Usage : $seq = $db->get_Seq_by_accession('AL12234')
Function: Gets a Bio::Seq object by its accession
Returns : a Bio::Seq object
Args : the id (as a string) of a sequence
Throws : "id does not exist" exception
NOTE: Bio::DB::RandomAccessI compliant method
get_Stream_by_acc ()
get_Seq_by_acc
Title : get_Seq_by_acc
Usage : $seq = $db->get_Seq_by_acc('X77802');
Function: Gets a Bio::Seq object by accession number
Returns : A Bio::Seq object
Args : accession number (as a string)
Throws : "acc does not exist" exception
NOTE: Bio::DB::RandomAccessI compliant method
get_Stream_by_id
Title : get_Stream_by_id
Usage : $seq = $db->get_Stream_by_id(@ids);
Function: Retrieves a stream of Seq objects given their ids
Returns : a Bio::SeqIO stream object
Args : an array of unique ids/accession numbers, or
an array reference
NOTE: This is also called get_Stream_by_batch()
get_Stream_by_batch ()
Title : get_Stream_by_batch
Usage : $seq = $db->get_Stream_by_batch(@ids);
Function: Retrieves a stream of Seq objects given their ids
Returns : a Bio::SeqIO stream object
Args : an array of unique ids/accession numbers, or
an array reference
NOTE: This is the same as get_Stream_by_id().
get_Stream_by_group ()
Bioperl compatibility.
all_seqfeatures
Title : all_seqfeatures
Usage : @features = $db->all_seqfeatures(@args)
Function: fetch all the features in the database
Returns : an array of features, or an iterator
Args : See below
Status : public
This is equivalent to calling $db->features() without any types, and will return all the features in the database. The -merge and -iterator arguments are recognized, and behave the same as described for features().
Creating and Loading GFF Databases
initialize
Title : initialize
Usage : $db->initialize(-erase=>$erase,-option1=>value1,-option2=>value2);
Function: initialize a GFF database
Returns : true if initialization successful
Args : a set of named parameters
Status : Public
This method can be used to initialize an empty database. It takes the following named arguments:
-erase A boolean value. If true the database will be wiped clean if it
already contains data.
Other named arguments may be recognized by subclasses. They become database meta values that control various settable options.
As a shortcut (and for backward compatibility) a single true argument is the same as initialize(-erase=>1).
load_gff
Title : load_gff
Usage : $db->load_gff($file|$directory|$filehandle);
Function: load GFF data into database
Returns : count of records loaded
Args : a directory, a file, a list of files,
or a filehandle
Status : Public
This method takes a single overloaded argument, which can be any of:
- 1. a scalar corresponding to a GFF file on the system
-
A pathname to a local GFF file. Any files ending with the .gz, .Z, or .bz2 suffixes will be transparently decompressed with the appropriate command-line utility.
- 2. an array reference containing a list of GFF files on the system
-
For example ['/home/gff/gff1.gz','/home/gff/gff2.gz']
- 3. directory path
-
The indicated directory will be searched for all files ending in the suffixes .gff, .gff.gz, .gff.Z or .gff.bz2.
- 4. filehandle
-
An open filehandle from which to read the GFF data.
- 5. a pipe expression
-
A pipe expression will also work. For example, a GFF file on a remote web server can be loaded with an expression like this:
$db->load_gff("lynx -dump -source http://stein.cshl.org/gff_test |");
If successful, the method will return the number of GFF lines successfully loaded.
NOTE:this method used to be called load(), but has been changed. The old method name is also recognized.
load_fasta
Title : load_fasta
Usage : $db->load_fasta($file|$directory|$filehandle);
Function: load FASTA data into database
Returns : count of records loaded
Args : a directory, a file, a list of files,
or a filehandle
Status : Public
This method takes a single overloaded argument, which can be any of:
- 1. scalar corresponding to a FASTA file on the system
-
A pathname to a local FASTA file. Any files ending with the .gz, .Z, or .bz2 suffixes will be transparently decompressed with the appropriate command-line utility.
- 2. array reference containing a list of FASTA files on the system
-
For example ['/home/fasta/genomic.fa.gz','/home/fasta/genomic.fa.gz']
- 3. path to a directory
-
The indicated directory will be searched for all files ending in the suffixes .fa, .fa.gz, .fa.Z or .fa.bz2.
a=item 4. filehandle
An open filehandle from which to read the FASTA data.
- 5. pipe expression
-
A pipe expression will also work. For example, a FASTA file on a remote web server can be loaded with an expression like this:
$db->load_gff("lynx -dump -source http://stein.cshl.org/fasta_test.fa |");
load_sequence_string
Title : load_sequence_string
Usage : $db->load_sequence_string($id,$dna)
Function: load a single DNA entry
Returns : true if successfully loaded
Args : a raw sequence string (DNA, RNA, protein)
Status : Public
lock_on_load
Title : lock_on_load
Usage : $lock = $db->lock_on_load([$lock])
Function: set write locking during load
Returns : current value of lock-on-load flag
Args : new value of lock-on-load-flag
Status : Public
This method is honored by some of the adaptors. If the value is true, the tables used by the GFF modules will be locked for writing during loads and inaccessible to other processes.
meta
Title : meta
Usage : $value = $db->meta($name [,$newval])
Function: get or set a meta variable
Returns : a string
Args : meta variable name and optionally value
Status : abstract
Get or set a named metavalues for the database. Metavalues can be used for database-specific settings.
By default, this method does nothing!
default_meta_values
Title : default_meta_values
Usage : %values = $db->default_meta_values
Function: empty the database
Returns : a list of tag=>value pairs
Args : none
Status : protected
This method returns a list of tag=>value pairs that contain default meta information about the database. It is invoked by initialize() to write out the default meta values. The base class version returns an empty list.
For things to work properly, meta value names must be UPPERCASE.
error
Title : error
Usage : $db->error( [$new error] );
Function: read or set error message
Returns : error message
Args : an optional argument to set the error message
Status : Public
This method can be used to retrieve the last error message. Errors are not reset to empty by successful calls, so contents are only valid immediately after an error condition has been detected.
debug
Title : debug
Usage : $db->debug( [$flag] );
Function: read or set debug flag
Returns : current value of debug flag
Args : new debug flag (optional)
Status : Public
This method can be used to turn on debug messages. The exact nature of those messages depends on the adaptor in use.
automerge
Title : automerge
Usage : $db->automerge( [$new automerge] );
Function: get or set automerge value
Returns : current value (boolean)
Args : an optional argument to set the automerge value
Status : Public
By default, this module will use the aggregators to merge groups into single composite objects. This default can be changed to false by calling automerge(0).
attributes
Title : attributes
Usage : @attributes = $db->attributes($id,$name)
Function: get the "attributres" on a particular feature
Returns : an array of string
Args : feature ID
Status : public
Some GFF version 2 files use the groups column to store a series of attribute/value pairs. In this interpretation of GFF, the first such pair is treated as the primary group for the feature; subsequent pairs are treated as attributes. Two attributes have special meaning: "Note" is for backward compatibility and is used for unstructured text remarks. "Alias" is considered as a synonym for the feature name.
If no name is provided, then attributes() returns a flattened hash, of attribute=>value pairs. This lets you do:
%attributes = $db->attributes($id);
Normally, attributes() will be called by the feature:
@notes = $feature->attributes('Note');
In a scalar context, attributes() returns the first value of the attribute if a tag is present, otherwise a hash reference in which the keys are attribute names and the values are anonymous arrays containing the values.
fast_queries
Title : fast_queries
Usage : $flag = $db->fast_queries([$flag])
Function: turn on and off the "fast queries" option
Returns : a boolean
Args : a boolean flag (optional)
Status : public
The mysql database driver (and possibly others) support a "fast" query mode that caches results on the server side. This makes queries come back faster, particularly when creating iterators. The downside is that while iterating, new queries will die with a "command synch" error. This method turns the feature on and off.
For databases that do not support a fast query, this method has no effect.
add_aggregator
Title : add_aggregator
Usage : $db->add_aggregator($aggregator)
Function: add an aggregator to the list
Returns : nothing
Args : an aggregator
Status : public
This method will append an aggregator to the end of the list of registered aggregators.
aggregators
Title : aggregators
Usage : $db->aggregators([@new_aggregators]);
Function: retrieve list of aggregators
Returns : list of aggregators
Args : a list of aggregators to set (optional)
Status : public
This method will get or set the list of aggregators assigned to the database. If 1 or more arguments are passed, the existing set will be cleared.
clear_aggregators
Title : clear_aggregators
Usage : $db->clear_aggregators
Function: clears list of aggregators
Returns : nothing
Args : none
Status : public
This method will clear the aggregators stored in the database object. Use aggregators() or add_aggregator() to add some back.
Methods for use by Subclasses
The following methods are chiefly of interest to subclasses and are not intended for use by end programmers.
abscoords
Title : abscoords
Usage : $db->abscoords($name,$class,$refseq)
Function: finds position of a landmark in reference coordinates
Returns : ($ref,$class,$start,$stop,$strand)
Args : name and class of landmark
Status : public
This method is called by Bio::DB::GFF::RelSegment to obtain the absolute coordinates of a sequence landmark. The arguments are the name and class of the landmark. If successful, abscoords() returns the ID of the reference sequence, its class, its start and stop positions, and the orientation of the reference sequence's coordinate system ("+" for forward strand, "-" for reverse strand).
If $refseq is present in the argument list, it forces the query to search for the landmark in a particular reference sequence.
Protected API
The following methods are not intended for public consumption, but are intended to be overridden/implemented by adaptors.
default_aggregators
Title : default_aggregators
Usage : $db->default_aggregators;
Function: retrieve list of aggregators
Returns : array reference containing list of aggregator names
Args : none
Status : protected
This method (which is intended to be overridden by adaptors) returns a list of standard aggregators to be applied when no aggregators are specified in the constructor.
do_load_gff
Title : do_load_gff
Usage : $db->do_load_gff
Function: load a GFF input stream
Returns : number of features loaded
Args : none
Status : protected
This method is called to load a GFF data stream. The method will read GFF features from <> and load them into the database. On exit the method must return the number of features loaded.
Note that the method is responsible for parsing the GFF lines. This is to allow for differences in the interpretation of the "group" field, which are legion.
setup_load
Title : setup_load
Usage : $db->setup_load
Function: called before load_gff_line()
Returns : void
Args : none
Status : abstract
This abstract method gives subclasses a chance to do any schema-specific initialization prior to loading a set of GFF records. It must be implemented by a subclass.
finish_load
Title : finish_load
Usage : $db->finish_load
Function: called after load_gff_line()
Returns : number of records loaded
Args : none
Status :abstract
This method gives subclasses a chance to do any schema-specific cleanup after loading a set of GFF records.
load_gff_line
Title : load_gff_line
Usage : $db->load_gff_line(@args)
Function: called to load one parsed line of GFF
Returns : true if successfully inserted
Args : see below
Status : abstract
This abstract method is called once per line of the GFF and passed a hashref containing parsed GFF fields. The fields are:
{ref => $ref,
class => $class,
source => $source,
method => $method,
start => $start,
stop => $stop,
score => $score,
strand => $strand,
phase => $phase,
gclass => $gclass,
gname => $gname,
tstart => $tstart,
tstop => $tstop,
attributes => $attributes}
do_initialize
Title : do_initialize
Usage : $db->do_initialize([$erase])
Function: initialize and possibly erase database
Returns : true if successful
Args : optional erase flag
Status : protected
This method implements the initialize() method described above, and takes the same arguments.
dna
Title : dna
Usage : $db->dna($id,$start,$stop,$class)
Function: return the raw DNA string for a segment
Returns : a raw DNA string
Args : id of the sequence, its class, start and stop positions
Status : public
This method is invoked by Bio::DB::GFF::Segment to fetch the raw DNA sequence.
Arguments: -name sequence name -start start position -stop stop position -class sequence class
If start and stop are both undef, then the entire DNA is retrieved. So to fetch the whole dna, call like this:
$db->dna($name_of_sequence);
or like this:
$db->dna(-name=>$name_of_sequence,-class=>$class_of_sequence);
NOTE: you will probably prefer to create a Segment and then invoke its dna() method.
get_dna
Title : get_dna
Usage : $db->get_dna($id,$start,$stop,$class)
Function: get DNA for indicated segment
Returns : the dna string
Args : sequence ID, start, stop and class
Status : protected
If start > stop and the sequence is nucleotide, then this method should return the reverse complement. The sequence class may be ignored by those databases that do not recognize different object types.
get_features
Title : get_features
Usage : $db->get_features($search,$options,$callback)
Function: get list of features for a region
Returns : count of number of features retrieved
Args : see below
Status : protected
The first argument is a hash reference containing search criteria for retrieving features. It contains the following keys:
rangetype One of "overlaps", "contains" or "contained_in". Indicates
the type of range query requested.
refseq ID of the landmark that establishes the absolute
coordinate system.
refclass Class of this landmark. Can be ignored by implementations
that don't recognize such distinctions.
start Start of the range, inclusive.
stop Stop of the range, inclusive.
types Array reference containing the list of annotation types
to fetch from the database. Each annotation type is an
array reference consisting of [source,method].
The second argument is a hash reference containing certain options that affect the way information is retrieved:
sort_by_group
A flag. If true, means that the returned features should be
sorted by the group that they're in.
sparse A flag. If true, means that the expected density of the
features is such that it will be more efficient to search
by type rather than by range.
The third argument, the $callback, is a code reference to which retrieved features are passed. It is described in more detail below.
This routine is responsible for getting arrays of GFF data out of the database and passing them to the callback subroutine. The callback does the work of constructing a Bio::DB::GFF::Feature object out of that data. The callback expects a list of 13 fields:
$refseq The reference sequence
$start feature start
$stop feature stop
$source feature source
$method feature method
$score feature score
$strand feature strand
$phase feature phase
$groupclass group class (may be undef)
$groupname group ID (may be undef)
$tstart target start for similarity hits (may be undef)
$tstop target stop for similarity hits (may be undef)
$feature_id A unique feature ID (may be undef)
These fields are in the same order as the raw GFF file, with the exception that the group column has been parsed into group class and group name fields.
The feature ID, if provided, is a unique identifier of the feature line. The module does not depend on this ID in any way, but it is available via Bio::DB::GFF->id() if wanted. In the dbi::mysql and dbi::mysqlopt adaptor, the ID is a unique row ID. In the acedb adaptor it is not used.
_feature_by_name
Title : _feature_by_name
Usage : $db->_feature_by_name($name,$class,$callback)
Function: get a list of features by name and class
Returns : count of number of features retrieved
Args : name of feature, class of feature, and a callback
Status : abstract
This method is used internally. The callback arguments are the same as those used by make_feature(). This method must be overidden by subclasses.
_feature_by_id
Title : _feature_by_id
Usage : $db->_feature_by_id($ids,$type,$callback)
Function: get a feature based
Returns : count of number of features retrieved
Args : arrayref to feature IDs to fetch
Status : abstract
This method is used internally to fetch features either by their ID or their group ID. $ids is a arrayref containing a list of IDs, $type is one of "feature" or "group", and $callback is a callback. The callback arguments are the same as those used by make_feature(). This method must be overidden by subclasses.
overlapping_features
Title : overlapping_features
Usage : $db->overlapping_features(@args)
Function: get features that overlap the indicated range
Returns : a list of Bio::DB::GFF::Feature objects
Args : see below
Status : public
This method is invoked by Bio::DB::GFF::Segment->features() to find the list of features that overlap a given range. It is generally preferable to create the Segment first, and then fetch the features.
This method takes set of named arguments:
-refseq ID of the reference sequence
-class Class of the reference sequence
-start Start of the desired range in refseq coordinates
-stop Stop of the desired range in refseq coordinates
-types List of feature types to return. Argument is an array
reference containing strings of the format "method:source"
-parent A parent Bio::DB::GFF::Segment object, used to create
relative coordinates in the generated features.
-rare Turn on an optimization suitable for a relatively rare feature type,
where it will be faster to filter by feature type first
and then by position, rather than vice versa.
-merge Whether to apply aggregators to the generated features.
-iterator Whether to return an iterator across the features.
If -iterator is true, then the method returns a single scalar value consisting of a Bio::SeqIO object. You can call next_seq() repeatedly on this object to fetch each of the features in turn. If iterator is false or absent, then all the features are returned as a list.
Currently aggregation is disabled when iterating over a series of features.
Types are indicated using the nomenclature "method:source". Either of these fields can be omitted, in which case a wildcard is used for the missing field. Type names without the colon (e.g. "exon") are interpreted as the method name and a source wild card. Regular expressions are allowed in either field, as in: "similarity:BLAST.*".
contained_features
Title : contained_features
Usage : $db->contained_features(@args)
Function: get features that are contained within the indicated range
Returns : a list of Bio::DB::GFF::Feature objects
Args : see overlapping_features()
Status : public
This call is similar to overlapping_features(), except that it only retrieves features whose end points are completely contained within the specified range.
Generally you will want to fetch a Bio::DB::GFF::Segment object and call its contained_features() method rather than call this directly.
contained_in
Title : contained_in
Usage : @features = $s->contained_in(@args)
Function: get features that contain this segment
Returns : a list of Bio::DB::GFF::Feature objects
Args : see features()
Status : Public
This is identical in behavior to features() except that it returns only those features that completely contain the segment.
get_abscoords
Title : get_abscoords
Usage : $db->get_abscoords($name,$class,$refseq)
Function: get the absolute coordinates of sequence with name & class
Returns : ($absref,$absstart,$absstop,$absstrand)
Args : name and class of the landmark
Status : protected
Given the name and class of a genomic landmark, this function returns a four-element array consisting of:
$absref the ID of the reference sequence that contains this landmark
$absstart the position at which the landmark starts
$absstop the position at which the landmark stops
$absstrand the strand of the landmark, relative to the reference sequence
If $refseq is provided, the function searches only within the specified reference sequence.
get_types
Title : get_types
Usage : $db->get_types($absref,$class,$start,$stop,$count)
Function: get list of all feature types on the indicated segment
Returns : list or hash of Bio::DB::GFF::Typename objects
Args : see below
Status : protected
Arguments are:
$absref the ID of the reference sequence
$class the class of the reference sequence
$start the position to start counting
$stop the position to end counting
$count a boolean indicating whether to count the number
of occurrences of each feature type
If $count is true, then a hash is returned. The keys of the hash are feature type names in the format "method:source" and the values are the number of times a feature of this type overlaps the indicated segment. Otherwise, the call returns a set of Bio::DB::GFF::Typename objects. If $start or $stop are undef, then all features on the indicated segment are enumerated. If $absref is undef, then the call returns all feature types in the database.
make_feature
Title : make_feature
Usage : $db->make_feature(@args)
Function: Create a Bio::DB::GFF::Feature object from string data
Returns : a Bio::DB::GFF::Feature object
Args : see below
Status : internal
This takes 14 arguments (really!):
$parent A Bio::DB::GFF::RelSegment object
$group_hash A hashref containing unique list of GFF groups
$refname The name of the reference sequence for this feature
$refclass The class of the reference sequence for this feature
$start Start of feature
$stop Stop of feature
$source Feature source field
$method Feature method field
$score Feature score field
$strand Feature strand
$phase Feature phase
$group_class Class of feature group
$group_name Name of feature group
$tstart For homologies, start of hit on target
$tstop Stop of hit on target
The $parent argument, if present, is used to establish relative coordinates in the resulting Bio::DB::Feature object. This allows one feature to generate a list of other features that are relative to its coordinate system (for example, finding the coordinates of the second exon relative to the coordinates of the first).
The $group_hash allows the group_class/group_name strings to be turned into rich database objects via the make_obect() method (see above). Because these objects may be expensive to create, $group_hash is used to uniquefy them. The index of this hash is the composite key {$group_class,$group_name,$tstart,$tstop}. Values are whatever object is returned by the make_object() method.
The remainder of the fields are taken from the GFF line, with the exception that "Target" features, which contain information about the target of a homology search, are parsed into their components.
parse_types
Title : parse_types
Usage : $db->parse_types(@args)
Function: parses list of types
Returns : an array ref containing ['method','source'] pairs
Args : a list of types in 'method:source' form
Status : internal
This method takes an array of type names in the format "method:source" and returns an array reference of ['method','source'] pairs. It will also accept a single argument consisting of an array reference with the list of type names.
make_match_sub
Title : make_match_sub
Usage : $db->make_match_sub($types)
Function: creates a subroutine used for filtering features
Returns : a code reference
Args : a list of parsed type names
Status : protected
This method is used internally to generate a code subroutine that will accept or reject a feature based on its method and source. It takes an array of parsed type names in the format returned by parse_types(), and generates an anonymous subroutine. The subroutine takes a single Bio::DB::GFF::Feature object and returns true if the feature matches one of the desired feature types, and false otherwise.
make_object
Title : make_object
Usage : $db->make_object($class,$name,$start,$stop)
Function: creates a feature object
Returns : a feature object
Args : see below
Status : protected
This method is called to make an object from the GFF "group" field. By default, all Target groups are turned into Bio::DB::GFF::Homol objects, and everything else becomes a Bio::DB::GFF::Featname. However, adaptors are free to override this method to generate more interesting objects, such as true BioPerl objects, or Acedb objects.
Arguments are:
$name database ID for object
$class class of object
$start for similarities, start of match inside object
$stop for similarities, stop of match inside object
do_attributes
Title : do_attributes
Usage : $db->do_attributes($id [,$tag]);
Function: internal method to retrieve attributes given an id and tag
Returns : a list of Bio::DB::GFF::Feature objects
Args : a feature id and a attribute tag (optional)
Status : protected
This method is overridden by subclasses in order to return a list of attributes. If called with a tag, returns the value of attributes of that tag type. If called without a tag, returns a flattened array of (tag=>value) pairs. A particular tag can be present multiple times.
Internal Methods
The following methods are internal to Bio::DB::GFF and are not guaranteed to remain the same.
_features
Title : _features
Usage : $db->_features($search,$options,$parent)
Function: internal method
Returns : a list of Bio::DB::GFF::Feature objects
Args : see below
Status : internal
This is an internal method that is called by overlapping_features(), contained_features() and features() to create features based on a parent segment's coordinate system. It takes three arguments, a search options hashref, an options hashref, and a parent segment.
The search hashref contains the following keys:
rangetype One of "overlaps", "contains" or "contained_in". Indicates
the type of range query requested.
refseq reference sequence ID
refclass reference sequence class
start start of range
stop stop of range
types arrayref containing list of types in "method:source" form
The options hashref contains zero or more of the following keys:
sparse turn on optimizations for a rare feature
automerge if true, invoke aggregators to merge features
iterator if true, return an iterator
The $parent argument is a scalar object containing a Bio::DB::GFF::RelSegment object or descendent.
get_features_iterator
Title : get_features_iterator
Usage : $db->get_features_iterator($search,$options,$callback)
Function: get an iterator on a features query
Returns : a Bio::SeqIO object
Args : as per get_features()
Status : Public
This method takes the same arguments as get_features(), but returns an iterator that can be used to fetch features sequentially, as per Bio::SeqIO.
Internally, this method is simply a front end to range_query(). The latter method constructs and executes the query, returning a statement handle. This routine passes the statement handle to the constructor for the iterator, along with the callback.
_split_group
Title : _split_group
Usage : $db->_split_group(@groups)
Function: parse GFF group field
Returns : ($gclass,$gname,$tstart,$tstop,$attributes)
Args : a list of group fields from a GFF line
Status : internal
This is an internal method that is called by load_gff_line to parse out the contents of one or more group fields. It returns the class of the group, its name, the start and stop of the target, if any, and an array reference containing any attributes that were stuck into the group field, in [attribute_name,attribute_value] format.
BUGS
Features can only belong to a single group at a time. This must be addressed soon.
Start coordinate can be greater than stop coordinate for relative addressing. This breaks strict BioPerl compatibility and must be fixed.
SEE ALSO
bioperl, Bio::DB::GFF::RelSegment, Bio::DB::GFF::Feature, Bio::DB::GFF::Adaptor::dbi::mysql, Bio::DB::GFF::Adaptor::dbi::mysqlopt
AUTHOR
Lincoln Stein <lstein@cshl.org>.
Copyright (c) 2001 Cold Spring Harbor Laboratory.
This library is free software; you can redistribute it and/or modify it under the same terms as Perl itself.