Orthologs finder

This image includes two commands to retrieve orthologous genes from input lists containing gene identifiers. The two commands are:

• diopt-orthologs: to obtain orthologs using the DIOPT Ortholog Finder. It allows using several gene identifiers as input, the same that appear in the 3. Enter Gene list combobox at the DIOPT online tool (default is Entrez GeneID).
• ensembl-orthologs: to obtain orthologs using the Ensembl REST API. It only allows using NCBI Gene IDs as input.

Both commands accept as input a TXT containing a list of Gene IDs (--gene_list_file), the input species (--input_species) and the target or output species (--output_species). Examples of both commands are provided below.

An additional command (list-species) is provided to show the taxonomy identifiers of the most common species: docker run --rm pegi3s/orthologs-finder list-species. This command can receive a parameter (uniprot_with_proteome or uniprot_with_kb_entries) to list taxonomy ID lists for species available at UniProt (it just shows pre-downloaded files obtained from UniProt taxonomy).

diopt-orthologs

The diopt-orthologs command also accepts these three parameters:

• -gs/--gene_list_search_fields=<search_fields>: the gene identifiers type. It must be one of the values that appear in the 3. Enter Gene list combobox at the DIOPT online tool (default is Entrez GeneID).
• -sd/--search_datasets=<search_datasets>: the ortholog sources. It must be one of the values that appear in the 4. Choose Ortholog Sources combobox at the DIOPT online tool (default is All).
• -af/--additional_filter=<additional_filter>: one of None, Best (return only best match when there is more than one match per input gene or protein), NoLow (default, exclude low score [score > 1, unless only match score is 1]), AboveTwo (exclude low scores [score > 2, unless only match score is 1 or 2]), or HighRank (exclude Low Ranked Scores).

You should adapt and run the following command to use it: docker run --rm -it -v "/your/data/dir:/data" pegi3s/orthologs-finder diopt-orthologs --input_species=<input_species> --output_species=<output_species> --gene_list_file=/path/to/gene_list_file --output=/path/to/output

Where you should replace:

• /your/data/dir to point to the directory that contains your input data.
• <input_species> and <output_species> with the taxonomy IDs of the input and output species (you can list the most common ones with docker run --rm pegi3s/orthologs-finder list-species).
• /path/to/gene_list_file to point to the path to the list of input gene identifiers.
• /path/to/output to point to the path where the output files will be created.

This script produces two TSV files:

• /path/to/output.tsv: the full conversion report provided by DIOPT.
• /path/to/output.mapping.tsv: a two-column TSV containing the gene identifiers in the input and target species.
• /path/to/output.unmapped.txt: a plain text file containing the gene identifiers in the input species that could not be mapped.

You can use the input files in the test_data directory to try this command.

docker run --rm -it -v "$(pwd)/test_data:/data" -w /data pegi3s/orthologs-finder diopt-orthologs --input_species=10090 --output_species=9606 --gene_list_file=gene_list_mus_musculus_10090 --output=gene_list_mus_musculus_10090_converted_to_Homo_sapiens.DIOPT

docker run --rm -it -v "$(pwd)/test_data:/data" -w /data pegi3s/orthologs-finder diopt-orthologs --input_species=7227 --output_species=9606 --gene_list_file=gene_list_dros_7227 --output=gene_list_dros_7227_converted_to_Homo_sapiens.DIOPT

docker run --rm -it -v "$(pwd)/test_data:/data" -w /data pegi3s/orthologs-finder diopt-orthologs --input_species=7955 --output_species=9606 --gene_list_file=gene_list_danio_rerio_7955 --output=gene_list_danio_rerio_7955_converted_to_Homo_sapiens.DIOPT

docker run --rm -it -v "$(pwd)/test_data:/data" -w /data pegi3s/orthologs-finder diopt-orthologs --input_species=9606 --output_species=6239 --gene_list_file=gene_list_homo_9606 --output=gene_list_homo_9606_converted_to_C_Elegans.DIOPT

docker run --rm -it -v "$(pwd)/test_data:/data" -w /data pegi3s/orthologs-finder diopt-orthologs --input_species=9606 --output_species=7227 --gene_list_file=gene_list_homo_9606 --output=gene_list_homo_9606_converted_to_Drosophila.DIOPT

docker run --rm -it -v "$(pwd)/test_data:/data" -w /data pegi3s/orthologs-finder diopt-orthologs --input_species=9606 --output_species=10090 --gene_list_file=gene_list_homo_9606 --output=gene_list_homo_9606_converted_to_Mus_musculus.DIOPT

docker run --rm -it -v "$(pwd)/test_data:/data" -w /data pegi3s/orthologs-finder diopt-orthologs --input_species=9606 --output_species=7955 --gene_list_file=gene_list_homo_9606 --output=gene_list_homo_9606_converted_to_Danio_rerio.DIOPT

Debugging

The script produces several temporary and intermediate files to create the final output. In case you obtain unexected results, you may want to inspect these temporary folder. To do so, just add -v /tmp:/tmp to the corresponding docker run command.

ensembl-orthologs

The ensembl-orthologs has no additional parameters appart from the input and output species taxonomy IDs and the input list of gene identifiers. Note that this script requires the gene identifiers to be NCBI Gene IDs. In case you need to map your identifiers, you may use the UniProt ID mapping tool or some other tool (e.g. Unipressed (Uniprot REST)).

You should adapt and run the following command to use it: docker run --rm -it -v "/your/data/dir:/data" pegi3s/orthologs-finder ensembl-orthologs --input_species=<input_species> --output_species=<output_species> --gene_list_file=/path/to/gene_list_file --output=/path/to/output

Where you should replace:

• /your/data/dir to point to the directory that contains your input data.
• <input_species> and <output_species> with the taxonomy IDs of the input and output species (you can list the most common ones with docker run --rm pegi3s/orthologs-finder list-species).
• /path/to/gene_list_file to point to the path to the list of input gene identifiers (NCBI Gene IDs).
• /path/to/output to point to the path where the output files will be created.

This script produces two TSV files:

• /path/to/output.tsv: the full conversion report provided by Ensembl. The first and second columns are the input gene identifier (source_gene_id and source_ensenbl_id) and the remaining ones are referred to the corresponding orthologous genes in the target species.
• /path/to/output.mapping.tsv: a two-column TSV containing the gene identifiers in the input (source_gene_id) and target species (id). Note that the gene identifiers on the target species may belong to different databases depending on the target species (e.g. if the target species is Drosophila melanogaster, these may be FlyBase [FBgnXXXX] identifiers; if the target species is Caenorhabditis elegans, these may be WormBase IDs [WBGeneXXXXX]; and if the target species are Homo sapiens, Mus musculus, or Danio rerio, these may be Ensembl IDs [ENSGXXXXX, ENSMUSGXXXXX, or ENSDARGXXXXX]).
• /path/to/output.unmapped.txt: a plain text file containing the gene identifiers in the input species that could not be mapped.

You can use the input files in the test_data directory to try this command.

docker run --rm -it -v "$(pwd)/test_data:/data" -w /data pegi3s/orthologs-finder ensembl-orthologs --input_species=10090 --output_species=9606 --gene_list_file=gene_list_mus_musculus_10090 --output=gene_list_mus_musculus_10090_converted_to_Homo_sapiens.Ensembl

docker run --rm -it -v "$(pwd)/test_data:/data" -w /data pegi3s/orthologs-finder ensembl-orthologs --input_species=7227 --output_species=9606 --gene_list_file=gene_list_dros_7227 --output=gene_list_dros_7227_converted_to_Homo_sapiens.Ensembl

docker run --rm -it -v "$(pwd)/test_data:/data" -w /data pegi3s/orthologs-finder ensembl-orthologs --input_species=7955 --output_species=9606 --gene_list_file=gene_list_danio_rerio_7955 --output=gene_list_danio_rerio_7955_converted_to_Homo_sapiens.Ensembl

docker run --rm -it -v "$(pwd)/test_data:/data" -w /data pegi3s/orthologs-finder ensembl-orthologs --input_species=9606 --output_species=6239 --gene_list_file=gene_list_homo_9606 --output=gene_list_homo_9606_converted_to_C_Elegans.Ensembl

docker run --rm -it -v "$(pwd)/test_data:/data" -w /data pegi3s/orthologs-finder ensembl-orthologs --input_species=9606 --output_species=7227 --gene_list_file=gene_list_homo_9606 --output=gene_list_homo_9606_converted_to_Drosophila.Ensembl

docker run --rm -it -v "$(pwd)/test_data:/data" -w /data pegi3s/orthologs-finder ensembl-orthologs --input_species=9606 --output_species=10090 --gene_list_file=gene_list_homo_9606 --output=gene_list_homo_9606_converted_to_Mus_musculus.Ensembl

docker run --rm -it -v "$(pwd)/test_data:/data" -w /data pegi3s/orthologs-finder ensembl-orthologs --input_species=9606 --output_species=7955 --gene_list_file=gene_list_homo_9606 --output=gene_list_homo_9606_converted_to_Danio_rerio.Ensembl

Notes

• The script uses the input species name to perform the queries. The species name associated to input taxonomy ID is obtained using the list-species command. If not found, then the input taxonomy ID is used, which in some cases may not work.
• The script makes one query to the Ensembl REST API for each gene identifier to be converted, waiting 1 second between queries by default. To increase or decrease this number, set the DELAY_BETWEEN_QUERIES environment variable (add -e DELAY_BETWEEN_QUERIES=0.5 to the docker run command).
• The script shows the gene being queried every 100 genes. To change this value, set the SHOW_GENES_INFO_BATCH_SIZE environment variable (add -e SHOW_GENES_INFO_BATCH_SIZE=<new_value> to the docker run command).

For instance, the last example command will be:

docker run --rm -it -v "$(pwd)/test_data:/data" -w /data -e DELAY_BETWEEN_QUERIES=0.5 -e SHOW_GENES_INFO_BATCH_SIZE=1 pegi3s/orthologs-finder ensembl-orthologs --input_species=9606 --output_species=7955 --gene_list_file=gene_list_homo_9606 --output=gene_list_homo_9606_converted_to_Danio_rerio.Ensembl

Debugging

To show additional debugging info, just add -e SHOW_DEBUGGING_INFO=enabled to the docker run command.