Software For Analyzing Gene Annotation On Genomes
Entering edit mode
10.1 years ago
baconator ▴ 20

Hello everyone.

I am looking for a way to programmatically access gene annotations and process them.

I have conducted an alignment of assembled short reads to the genome of Pseudomonas fluorescens SBW25 (NC_012660.1). I have found a large gap and I would like to find out which genes may be located in that gap. I know the genomic coordinates of the start and end of the gap.

I have used the OperonDB to find a list of genes in that gap (see However, there are several dozen genes in the gap and I would like to conduct analyses on the entire set. Therefore, I would like to find a way to download a gene annotation and take the subset delimited by the start and end of my gap.

My preferred working environment is R. I also have facility with Perl and Java.

Any help would be greatly appreciated.

genome sequence gene annotation • 3.1k views
Entering edit mode
10.1 years ago

If you can convert your gene annotations into a BED file - say, converting FASTA-formatted annotations to BED with Galaxy or other tools - and your genomic range in a set of BED coordinates, then you can use the bedmap tool in the BEDOPS suite to calculate a result containing those genes located within your genomic range(s) of interest.

To demonstrate, let's say your gene annotations are formatted as a BED file called annotations.bed, like so:

chr1    1000    1500    gene-1
chr1    4000    4300    gene-2
chr1    6200    6450    gene-3
chr1    9120    9675    gene-4

We're following UCSC's definition of a BED file, with the first column representing the chromosome name, the second and third columns the genomic coordinates, and the fourth column the ID or name of the annotation.

Let's say you want to do an ad-hoc search through this file, to find genes on the chromosome chr1 which are contained within the range [2000, 7000). You could use bedmap as follows:

$ echo -e "chr1\t2000\t7000" | bedmap --echo --echo-map-id - annotations.bed
chr1    2000    7000|gene-2;gene-3

The result indicates that gene-2 and gene-3 are contained within the BED coordinate range [2000, 7000) on chromosome chr1.

The echo -e command passes bedmap that range via standard input, which bedmap consumes with the - hyphen character. The bedmap tool then maps annotations to that input range and shows you these results.

Results are a semi-colon-delimited list of genes for that range, taken from the ID column of annotations.bed. This is because we used the --echo-map-id operator, which outputs mapped IDs. There are four --echo-map-* operators that offer the entire mapped annotation, or various pieces of it (ID, genomic range, scores). Please refer to the documentation for more detail.

The output from a bedmap operation is trivial to parse with Perl scripts or Java programs, through the usual split() or regular expression calls, etc.

Instead of a trivial example that uses echo and standard input, you can instead specify a BED file containing multiple ranges, each on their own line. For example, let's say we have a file called rangesOfInterest.bed that contains two ranges (you can specify as many ranges as you like, each on their own line):

chr1    2000    7000
chr1    5000    10000

You can then use bedmap as follows to get results containing the annotations of interest:

$ bedmap --echo --echo-map-id rangesOfInterest.bed annotations.bed
chr1    2000    7000|gene-2;gene-3
chr1    5000    10000|gene-3;gene-4

Further, if you want BED-formatted output, use the --delim operator in conjunction with --echo, as follows:

$ bedmap --echo --echo-map-id --delim '\t' rangesOfInterest.bed annotations.bed

This replaces the pipe character (|) with a tab (\t), so that the output is a minimal BED-formatted result:

$ bedmap --echo --echo-map-id --delim '\t' rangesOfInterest.bed annotations.bed
chr1    2000    7000    gene-2;gene-3
chr1    5000    10000   gene-3;gene-4

This trick is very useful if you want to pipe the results to another BEDOPS command or another utility that consumes BED-formatted data, like awk, Perl or Python scripts, etc.

BEDOPS tools are modular and written to accept standard input (such as what was done above with echo -e), so it is easy to perform further set or mapping operations with bedops and bedmap, respectively, or compress the output with starch, etc. in an extended pipeline. Piping standard output from one program to standard input of another program reduces overall I/O load on a computer and can improve the speed of calculations.

The bedmap tool is also useful for quantitative work. While the gene annotations in this example do not contain any numerical data, we might want to quickly count how many genes are binned within our ranges of interest, using the --count operator:

$ bedmap --echo --echo-map-id --count --delim '\t' rangesOfInterest.bed annotations.bed
chr1    2000    7000    gene-2;gene-3    2
chr1    5000    10000   gene-3;gene-4    2

Granted, this is a pretty mundane example, but you could imagine asking how many annotations are contained within a moving window over a genome. This would tell you where your annotations are concentrated, which could have biological significance.

Other numerical operators are available for bedmap, useful particularly if your mapped elements contain score information, like tag counts, p-values, expression levels, etc.

To perform all these operations within R, use a system() call.


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