Hi!

I'm trying to use GOATOOLS to perform Gene Ontology Enrichment analyses on a list of gene from a non-model organism (Anopheles gambiae, main vector of human malaria).

I managed to run the tool, but I'm having issue with the statistical analyses since I'm not getting any output. I'm using a gaf file to build my reference dataset.

This is my script, it might have a lot of missing gap.

import numpy as np
from goatools.base import download_go_basic_obo
from goatools.base import download_ncbi_associations
from goatools.obo_parser import GODag
from goatools.associations import read_gaf
from goatools.anno.genetogo_reader import Gene2GoReader
from goatools.goea.go_enrichment_ns import GOEnrichmentStudy


#obo_fname = download_go_basic_obo()
obodag = GODag('GO_annotation/go-basic.obo')

gaf_file = ('GO_annotation/VectorBase-CURRENT_AgambiaePEST_GO.gaf')  # Path to your GAF file

geneid2gos = read_gaf(gaf_file, namespace='BP', godag=obodag)

geneid2gos

goeaobj = GOEnrichmentStudy(
    AGAP_YSHR_ZGA, # my list of genes
    geneid2gos, # List of anopheles gene
    obodag, #Ontologies
    propagate_counts = False,
    alpha = 0.05,
    methods= ['fdr_tsbh'])

#goeaobj
goea_results_all = goeaobj.run_study(AGAP_YSHR_ZGA)

goea_results_all

This is the output that I'm getting:

GO_annotation/go-basic.obo: fmt(1.2) rel(2023-05-10) 46,490 Terms
HMS:0:00:03.154457 120,533 annotations READ: GO_annotation/VectorBase-CURRENT_AgambiaePEST_GO.gaf 
7755 IDs in loaded association branch, BP

Load  Ontology Enrichment Analysis ...
 64%  5,229 of  8,162 population items found in association

Runing  Ontology Analysis: current study set of 8162 IDs.
 64%  5,229 of  8,162 study items found in association
100%  8,162 of  8,162 study items found in population(8162)
Calculating 2,439 uncorrected p-values using fisher_scipy_stats
   2,439 terms are associated with  5,229 of  8,162 population items
   2,439 terms are associated with  5,229 of  8,162 study items
  METHOD fdr_tsbh:
       0 GO terms found significant (< 0.05=alpha) (  0 enriched +   0 purified): statsmodels fdr_tsbh
       0 study items associated with significant GO IDs (enriched)
       0 study items associated with significant GO IDs (purified)

It's seems that is able to find my list fo gene in the dataset, but is not performing any statistical test. I know that some of the genes are enriched since I used PANTHER that gave me the right enrichment.

Can someone help me?

Thanks in advance!