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Build .gbk files starting from eggnog annotation files and genomes (fasta)

Project description

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emapper2gbk: creation of genbank files from Eggnog-mapper annotation outputs

Starting from fasta and Eggnog-mapper annotation files, build a gbk file that is suitable for metabolic network reconstruction with Pathway Tools. Adds the GO terms and EC numbers annotations in the genbank file. There are two main modes:

  • genes mode: suitable when a list of isolated genes/proteins have been annotated with Eggnog-mapper, typically the gene catalogue of a metagenome.
  • genomes mode: usually when focusing on a single organism, with a .gff file. The creation of genbanks can be performed in parallel by providing directories (with matching names for genomes, proteomes and annotation files) as inputs.

If you use emapper2gbk, please cite

Belcour* A, Frioux* C, Aite M, Bretaudeau A, Hildebrand F, Siegel A. Metage2Metabo, microbiota-scale metabolic complementarity for the identification of key species. eLife 2020;9:e61968 https://doi.org/10.7554/eLife.61968 .

Main inputs

emapper2gbk genes

For each annotated list of genes, inputs are:

  • a nucleotide fasta file containing the CDS sequence of each genes or a folder containing multiple nucleotide fasta files.
  • the translated sequences in amino-acids in fasta or a folder containing the corresponding protein sequences to the nucleotide sequences (must be the same name).
  • the annotation file obtained after Eggnog-mapper annotation (usually xxx.emapper.annotation) or a folder with multiple annotation files (must be the same name as nucleotide fasta file and ends with ‘.tsv’ extension).

In addition, as optional files:

  • the name of the considered organism (can be “bacteria” or “metagenome”) or a file with organisms names (matching the genomes names).
  • the merge option to merge genes into fake contigs.
  • the number of available cores for multiprocessing (when working on multiple genomes).
  • a go-basic file of GO ontology (if not given, emapper2gbk will download a copy and use it).

Example:

Input with files:

nucleotide_sequences.fna
protein_sequence.faa
annotation.emapper.annotation

Input with folders:

nucleotide_sequences
├── gene_list_1.fna
├── gene_list_2.fna
protein_sequence
├── gene_list_1.faa
├── gene_list_2.faa
annotation
├── gene_list_1.tsv
├── gene_list_2.tsv
pictures/emapper2gbk_genes.svg

To work the ID of the genes in the nucleic fasta file (-fn) must be the same than the ID of the proteins in the protein fasta file (-fp) and in the annotation file (-a).

emapper2gbk genomes

For each genomes, inputs are:

  • a nucleotide fasta file containing the sequence of each contigs/chromosomes for the genome or a folder containing multiple nucleotide fasta files.
  • the proteome corresponding to the genome or a folder containing the corresponding protein sequences to the nucleotide sequences (must be the same name).
  • the GFF file corresponding to the genome or a folder containing multiple GFF files (must be the same name as the nucleotide folder).
  • the annotation file obtained after Eggnog-mapper annotation (usually xxx.emapper.annotation) or a folder with multiple annotation files (must be the same name as nucleotide fasta file and ends with ‘.tsv’ extension)

In addition, as optional files:

  • the name of the considered organism (can be “bacteria”) or a file with organisms names (matching the genomes names).
  • the number of available cores for multiprocessing (when working on multiple genomes).
  • a go-basic file of GO ontology (if not given, emapper2gbk will download a copy and use it).

Example:

Input with files:

nucleotide_sequences.fna
protein_sequence.faa
annotation.emapper.annotation
genome.gff

Input with folders:

nucleotide_sequences
├── genome_1.fna
├── genome_2.fna
protein_sequence
├── genome_1.faa
├── genome_2.faa
annotation
├── genome_1.tsv
├── genome_2.tsv
gff
├── genome_1.gff
├── genome_2.gff
pictures/emapper2gbk_genomes.svg

The ID in the chromosome/contigs/scaffolds fasta file (-fn) must correspond to region in the gff file (-g). Then the genes in the region will be found and the child CDS associated to the genes wil be extracted. The CDS ID must be the same than the ID in the protein fasta file (-fp) and the ID in the eggnog-mapper annotation file (-a).

By default emapper2gbk searches for inheritance between genes and CDS in the GFF files. A gene feature is required and the CDS feature must have the gene feature as a parent, like in this example:

##gff file
region_1    RefSeq  region  1       12642   .       +       .       ID=region_1
region_1    RefSeq  gene    1       2445    .       -       .       ID=gene_1
region_1    RefSeq  CDS     1       2445    .       -       0       ID=cds_1;Parent=gene_1

But some GFF files can be formatted differently, so by using the argument -gt cds_only, it is possible to use GFF file with only CDS, like:

##gff file
region_1    RefSeq  region  1       12642   .       +       .       ID=region_1
region_1    RefSeq  CDS     1       2445    .       -       0       ID=cds_1

Dependencies and installation

Dependencies

All are described in requirements.txt and can be installed with pip install -r requirements.txt.

  • biopython
  • gffutils
  • pandas
  • pronto
  • requests

Install

  • From this cloned repository
pip install -r requirements.txt
pip install .
  • From Pypi
pip install emapper2gbk

Usage

Convert GFF, fastas, annotation table and species name into Genbank.

usage: emapper2gbk [-h] [-v] {genes,genomes} ...

Starting from fasta and Eggnog-mapper annotation files, build a gbk file that is suitable for metabolic network reconstruction with Pathway Tools. Adds the GO terms and EC numbers annotations in the genbank file.

Two modes:
- genomes (one genome/proteome/gff/annot file --> one gbk).
- genes with the annotation of the full gene catalogue and fasta files (nucleic and protein) corresponding to list of genes.

Examples:

* Genomic - single mode

emapper2gbk genomes -fn genome.fna -fp proteome.faa -gff genome.gff -n "Escherichia coli" -o coli.gbk -a eggnog_annotation.tsv [-go go-basic.obo]

* Genomic - multiple mode, "bacteria" as default name

emapper2gbk genes -fn genome_dir/ -fp proteome_dir/ -n metagenome -o gbk_dir/ -a eggnog_annotation_dir/ [-go go-basic.obo]

* Genomic - multiple mode, tsv file for organism names

emapper2gbk genes -fn genome_dir/ -fp proteome_dir/ -nf matching_genome_orgnames.tsv -o gbk_dir/ -a eggnog_annotation_dir/ [-go go-basic.obo]

* Metagenomic

emapper2gbk genes -fn genome_dir/ -fp proteome_dir/ -o gbk_dir/ -a gene_cat_ggnog_annotation.tsv --one-annot-file [-go go-basic.obo]

You can give the GO ontology as an input to the program, it will be otherwise downloaded during the run. You can download it here: http://purl.obolibrary.org/obo/go/go-basic.obo .
The program requests the NCBI database to retrieve taxonomic information of the organism. However, if the organism is "bacteria" or "metagenome", the taxonomic information will not have to be retrieved online.
Hence, if you need to run the program from a cluster with no internet access, it is possible for a "bacteria" or "metagenome" organism, and by providing the GO-basic.obo file.
For specific help on each subcommand use: emapper2gbk {cmd} --help

optional arguments:
-h, --help       show this help message and exit
-v, --version    show program's version number and exit

subcommands:
valid subcommands:

{genes,genomes}
    genes          genes mode : 1-n annot, 1-n faa, 1-n fna (gene sequences) --> 1 gbk
    genomes        genomes mode: 1-n contig/chromosome fasta, 1-n protein fasta, 1-n GFF, 1-n annot --> 1 gbk
  • Genomes mode

    • Usage

      usage: emapper2gbk genomes [-h] -fn FASTANUCLEIC -fp FASTAPROT -o OUPUT_DIR -g GFF [-gt GFF_TYPE] [-nf NAMEFILE]
                              [-n NAME] -a ANNOTATION [-c CPU] [-go GOBASIC] [-q] [--keep-gff-annotation]
      
      Build a gbk file for each genome with an annotation file for each
      
      optional arguments:
      -h, --help            show this help message and exit
      -fn FASTANUCLEIC, --fastanucleic FASTANUCLEIC
                              fna file or directory
      -fp FASTAPROT, --fastaprot FASTAPROT
                              faa file or directory
      -o OUPUT_DIR, --out OUPUT_DIR
                              output directory/file path
      -g GFF, --gff GFF     gff file or directory
      -gt GFF_TYPE, --gff-type GFF_TYPE
                              gff type, by default emapper2gbk search for CDS with gene as Parent in the GFF, but by using
                              the '-gt cds_only' option emapper2gbk will only use the CDS information from the genome
      -nf NAMEFILE, --namefile NAMEFILE
                              organism/genome name (col 2) associated to genome file basenames (col 1). Default =
                              'metagenome' for metagenomic and 'cellular organisms' for genomic
      -n NAME, --name NAME  organism/genome name in quotes
      -a ANNOTATION, --annotation ANNOTATION
                              eggnog annotation file or directory
      -c CPU, --cpu CPU     cpu number for metagenomic mode or genome mode using input directories
      -go GOBASIC, --gobasic GOBASIC
                              go ontology, GOBASIC is either the name of an existing file containing the GO Ontology or the
                              name of the file that will be created by emapper2gbk containing the GO Ontology
      -q, --quiet           quiet mode, only warning, errors logged into console
      --keep-gff-annotation
                              Copy the annotation from the GFF (product) into the genbank output file.
      
    • Examples

      • Genomic - single mode
      emapper2gbk genomes -fn genome.fna -fp proteome.faa -gff genome.gff -n "Escherichia coli" -o coli.gbk -a eggnog_annotation.tsv [-go go-basic.obo]
      
      • Genomic - multiple mode, “bacteria” as default name
  • genes mode

    • Usage

      usage: emapper2gbk genes [-h] -fn FASTANUCLEIC -fp FASTAPROT -o OUPUT_DIR [--one-annot-file] -a ANNOTATION [-c CPU]
                              [-n NAME] [-nf NAMEFILE] [-go GOBASIC] [--merge MERGE] [-q]
      
      Build a gbk file for each genome/set of genes with an annotation file for each
      
      optional arguments:
      -h, --help            show this help message and exit
      -fn FASTANUCLEIC, --fastanucleic FASTANUCLEIC
                              fna file or directory
      -fp FASTAPROT, --fastaprot FASTAPROT
                              faa file or directory
      -o OUPUT_DIR, --out OUPUT_DIR
                              output directory/file path
      --one-annot-file      Option to use when there is only one annotation file for multiples genes fastas.
      -a ANNOTATION, --annotation ANNOTATION
                              eggnog annotation file or directory
      -c CPU, --cpu CPU     cpu number for metagenomic mode or genome mode using input directories
      -n NAME, --name NAME  organism/genome name in quotes
      -nf NAMEFILE, --namefile NAMEFILE
                              organism/genome name (col 2) associated to genome file basenames (col 1). Default =
                              'metagenome' for metagenomic and 'cellular organisms' for genomic
      -go GOBASIC, --gobasic GOBASIC
                              go ontology, GOBASIC is either the name of an existing file containing the GO Ontology or the
                              name of the file that will be created by emapper2gbk containing the GO Ontology
      --merge MERGE         Number of gene sequences to merge into fake contig from a same file in the genbank file.
      -q, --quiet           quiet mode, only warning, errors logged into console
      
    • Example

      emapper2gbk genes -fn genome_dir/ -fp proteome_dir/ -o gbk_dir/ -a gene_cat_ggnog_annotation.tsv [-go go-basic.obo]
      

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