Molecular Characterization, Expression Profiling, and SNP Analysis of the Porcine RNF20 Gene

SIMPLE SUMMARY: In this study, we found that RNF20 is ubiquitously expressed in porcine tissues, and the sequence of the RING domain was highly conserved across different species. Eight potential single nucleotide polymorphisms (SNPs) were discovered, and one of them, SNP1 (A-1027G), was confirmed by PCR-restriction fragment length polymorphism (RFLP). Allele frequency differences were also analyzed in four pig breeds. This study provides a preliminary understanding of the porcine RNF20 gene. ABSTRACT: Fat deposition is considered an economically important trait in pig breeding programs. Ring finger protein 20 (RNF20), an E3 ubiquitin protein ligase, has been shown to be closely involved in adipogenesis in mice, suggesting its conserved role in pigs. In this study, we obtained the exon sequences of the porcine RNF20 gene and characterized its molecular sequence. The porcine RNF20 gene contains 20 exons that encode 975 amino acids, and its RING domain is highly conserved across different species. Western blot analysis revealed that RNF20 was widely expressed, especially in various fat depots, and the level of H2B monoubiquitination (H2Bub) was highly consistent. Eight potential SNPs were detected by sequencing pooled PCR fragments. PCR–RFLP was developed to detect a single nucleotide polymorphism (A-1027G) in exon 1, and the allele frequency differences were examined in four pig breeds. The G allele was predominant in these pigs. Association analysis between (A-1027G) and the backfat thickness of three commercial pig breeds was performed, but no significant association was found. Taken together, these results enabled us to undertake the molecular characterization, expression profiling, and SNP analysis of the porcine RNF20 gene.