Abstract:

The study aimed to explore the effect of overexpression of TRIM39.2, localized in the chicken major histocompatibility complex (MHC), on the transcription expression of chicken macrophages. The tissue expression profile of chicken TRIM39.2 gene was constructed using RT-qPCR.The chicken TRIM39.2 gene was cloned and a recombinant expression vector carrying the tag was constructed. HD11 cells were transfected with either an empty vector or a TRIM39.2 eukaryotic expression vector.RNA was harvested to prepare a library, and transcriptome sequencing and bioinformatics analysis were performed. Differentially expressed genes were screened, followed by GO and KEGG enrichment analysis, and RT-qPCR method was used to further verify the transcriptome sequencing results. The results showed that the expression of TRIM39.2 was the highest in the spleen compared to other tissues. The chicken TRIM39.2 gene was successfully cloned, and a recombinant expression vector carrying a Myc tag, pCAGGS-Myc-TRIM39.2, was constructed, and confirmed to express TRIM39.2 recombinant protein in chicken HD11 cells.Transcriptome sequencing analysis identified 33 significantly differentially expressed genes. GO analysis was primarily enriched in the active characteristics of cytokines and their involvement in signal transduction pathways.KEGG analysis mainly focused on the interactions between cytokines and receptors, as well as pathways such as RIG-I-like receptor signaling pathway. The chicken TRIM39.2 gene plays a significant regulatory role in macrophage-mediated immune responses and signal transduction, particularly in positively regulating the type Ⅰ interferon signaling pathway mediated by RIG-I-like receptors. This study lays the foundation for further exploration of the regulatory mechanism of RIG-I-like receptor signaling pathway in chickens.

Key words: TRIM39.2, chicken macrophages, transcriptome, GO, KEGG