Gage GO annotations vs Entrez
Hi all,
I'm using Gage v2.28.2 to analyse gene enrichment pathways in RNA-seq data. Genes were initially annotated with entrez ID, which worked fine:
kg.eco=kegg.gsets("eco")
kg.eco.eg=kegg.gsets("eco", id.type = "entrez")
headkg.eco.eg$kg.sets, 2)
keggres = gage(p3, gsets=kg.eco.eg$kg.sets, ref = ref.idx, samp = samp.idx, same.dir = F)
lapply(keggres, head)
$greater
p.geomean stat.mean
eco02010 ABC transporters 5.636397e-06 4.437748
eco01100 Metabolic pathways 4.192560e-07 3.560120
eco02020 Two-component system 2.976999e-03 2.635986
eco02060 Phosphotransferase system (PTS) 1.029505e-02 2.316506
eco02024 Quorum sensing 8.269080e-03 2.290890
eco01120 Microbial metabolism in diverse environments 8.855620e-04 2.186511
p.val q.val
eco02010 ABC transporters 1.255645e-09 6.654916e-08
eco01100 Metabolic pathways 4.372851e-07 1.158805e-05
eco02020 Two-component system 1.382499e-04 2.442415e-03
eco02060 Phosphotransferase system (PTS) 8.903017e-04 1.053680e-02
eco02024 Quorum sensing 9.940376e-04 1.053680e-02
eco01120 Microbial metabolism in diverse environments 1.307795e-03 1.076939e-02
Input data is the same except row names are changed to GO terms:
head(p3)
Bg_NB25_v_BgNS26_2h_fc Bg_NB25_v_BgNP27_2h_fc
<NA> 1.559100 1.492170
GO:0000150|GO:0003677|GO:0006310 1.696600 1.251170
<NA> 0.688138 0.403168
GO:0003824 0.770600 0.744205
GO:0006355 1.185640 1.403170
GO:0008982|GO:0009401|GO:0016020 2.092530 0.818206
Bg_NB31_v_BgNS32_2h_fc Bg_NB31_v_BgNP33_2h_fc
<NA> -0.0207885 0.401330
GO:0000150|GO:0003677|GO:0006310 1.3511800 0.285852
<NA> 0.3511800 -0.299110
Then I ran the following commands:
data(go.sets.hs)
data(go.subs.hs)
keggres = gage(p3, gsets=go.sets.hs[go.subs.hs$BP], same.dir = F)
The results contain only NA's, I'm not sure why this is?
lapply(keggres, head)
$greater
p.geomean stat.mean p.val q.val
GO:0000002 mitochondrial genome maintenance NA NaN NA NA
GO:0000003 reproduction NA NaN NA NA
GO:0000012 single strand break repair NA NaN NA NA
GO:0000018 regulation of DNA recombination NA NaN NA NA
GO:0000019 regulation of mitotic recombination NA NaN NA NA
GO:0000022 mitotic spindle elongation NA NaN NA NA
set.size Bg_NB31_v_BgNS32_2h_fc
GO:0000002 mitochondrial genome maintenance 0 NA
GO:0000003 reproduction 0 NA
GO:0000012 single strand break repair 0 NA
GO:0000018 regulation of DNA recombination 0 NA
GO:0000019 regulation of mitotic recombination 0 NA
GO:0000022 mitotic spindle elongation 0 NA
There are a total of ~13,0000 genes in the original mapping database. Only ~32000 of these have GO annotations. The vast majority of enriched genes should be bacterial.
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Remove the
NArow names from thep3object and try again.Thanks for the reply. Same result with NA row names removed: