猪NR1H3基因可变剪接体的克隆及表达特性研究

doi:10.11843/j.issn.0366-6964.2020.01.006

Abstract

Abstract: The aim of this study was to clone the alternative splicing isoforms of pig NR1H3 gene, to predict the structures and functions of their coding proteins, and to investigate the expression patterns of each transcript. The Mashen pig was used in this study. The full-length coding sequence (CDS) of NR1H3 gene in Mashen pig was cloned and sequenced, the biological characteristics of NR1H3 protein was analyzed by bioinformatics methods, and the expression patterns of NR1H3 gene in heart, liver, spleen, lung, kidney, stomach, small intestine, biceps femoris, psoas magnus and subcutaneous fat and the chronological expression trends in adipose tissue in Mashen pig were detected by quantitative real-time PCR (qRT-PCR). The results showed that two of the transcripts of NR1H3 gene, NR1H3-1 and NR1H3-2, were successfully obtained. NR1H3-2 was a new transcript (MN082630), and the length of CDS was 1 203 bp, which encoded 400 amino acids. The encoded protein was neutral unstable protein. Compared with the NR1H3-1 transcript, NR1H3-2 had a deletion of the 9th exon with a length of 95 bp. Coding protein NR1H3-2 had a deletion of ligand binding domain of liver X receptors compared with NR1H3-1. The similarity of NR1H3 amino acid sequence of pig to those of Balaenoptera acutorostrata scammoi, Capra hircus, Ovis aries, Physeter catodon, Neophocaena asiaeorientalis asiaeorientalis was higher. NR1H3-1 and NR1H3-2 were expressed in all detected tissues in this study, and the expression differences among different tissues were significant (P<0.05). The highest abundance of NR1H3-1 was in small intestine, followed by spleen, liver and biceps femoris, and the lowest expression level was in heart. The highest abundance of NR1H3-2 was in liver, followed by spleen, small intestine, stomach and biceps femoris, and the lowest expression level was in heart. The abundance of NR1H3-2 in all tissues except for small intestine, heart and lung were higher than that of NR1H3-1, and the expression difference between NR1H3-1 and NR1H3-2 was extremely significant in liver and subcutaneous adipose tissue (P<0.01), and the difference was significant in stomach (P<0.05). With the increase of age, the expressions of NR1H3-1 and NR1H3-2 in adipose tissue in Mashen pig showed an overall upward trend, which could be deduced that NR1H3 gene might play an important role in the regulation of fat deposition in pig. In this study, two transcripts of pig NR1H3 gene were successfully cloned. It was speculated that NR1H3 might play important physiological roles in lipid metabolism and fat deposition in pig.

Key words: pig, NR1H3 gene, alternative splicing, bioinformatics, expression patterns

CLC Number:

S828.2

ZHANG Xuelian, JIN Dapeng, WU Yiqi, LI Wenxia, QIN Benyuan, CAI Chunbo, GAO Pengfei, GUO Xiaohong, LI Bugao, CAO Guoqing. Molecular Cloning of Alternative Splice Variants of NR1H3 Gene and Their Expression Patterns in Pig[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(1): 43-54.

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Molecular Cloning of Alternative Splice Variants of NR1H3 Gene and Their Expression Patterns in Pig

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