(f) Bioinformatics analysis
To study the enriched biological function and significant pathways of the differentially expressed proteins, the identified proteins UniProt IDs were used to run Gene Ontology (GO) term analysis. The homology search was performed for all identified sequences against the zebra finch (Taeniopygia guttata ) using NCBI BLASTX program (Kent, 2002). The zebra finch Uniprot IDs were assigned for the best hit sequence for GO annotation and enrichment analysis using ShinyGO online software (Ge et al., 2020). The GO enrichment results were presented in three functional categories based on the properties of the identified proteins: biological process (BP), cellular component (CC), and molecular function (MF). In addition, all the differentially expressed proteins were searched against the Kyoto Encyclopedia of Gene and Genomes (KEGG) database to identify the significant KEGG pathways. To further identify significant mechanisms underlying differentially expressed proteins associated with functional clusters of DEPs, GO term enrichment and KEGG pathways analysis was performed by Fisher’s exact test. GO terms with FDR values < 0.05 and KEGG pathways with FDR p values < 0.1 were identified as significantly enriched.
Results