陆川猪黑皮质激素受体4基因克隆及序列分析

doi:10.16431/j.cnki.1671-7236.2018.02.005

Abstract

Abstract:

This study was aimed to clone melanocortin-4 receptor(MC4R) gene of Luchuan pig and analyze its genetic structure with bioinformatic software. The recombination plasmid pMD18-T-MC4R was constructed with RT-PCR and cloning methods, and detected by colony PCR and sequencing. After successfully constructed the recombination plasmid pMD18-T-MC4R, the further analysis of physical and chemical properties, structure of protein and modification and the subcellular localization were proceeded. The results showed that the cloned MC4R gene fragment included a 999 bp whole length CDS and encoded 332 amino acids. Compared with MC4R gene of Sus scrofa submission in NCBI, there were four base mutations of MC4R gene in Luchuan pig, two mutations at 175 and 906 bp were the same sense mutations,the other two mutations at 110 and 278 bp were the mis-sense mutations, causing glutamic acid converted to glycine and valine converted to alanine in 37 and 93 amino acid residues, respectively. Homology analysis indicated that MC4R was evolutionarily conserved in different species. Seven transmembrane helical regions, several N-glycosylation sites and distinct hydrophobic region were found, but no signal peptide and potential phosphorylation exited in MC4R protein, which might distribute mainly in the endoplasmic reticulum and vacuolar. The elements in the second structure of encoded protein were α-helix, extended strand,β-turn and random coil. The Luchuan pig MC4R gene was successfully cloned in this study, and provided a foundation for further studying on developing and utilizing the local species Luchuan pig and its breeding.

Key words: Luchuan pig; MC4R gene; clone

CLC Number:

XIE Wan, LIU Mingjun, HE Jianxiong, SHEN Yujian, CHEN Baojian, GUAN Zhihui, ZHANG Mingyuan, GUO Yafen, LAN Ganqiu, JIANG Qinyang, YANG Xiurong, LIANG Jing, XIE Bingkun. Cloning and Sequence Analysis of MC4R Gene in Luchuan Pig[J]. , 2018, 45(2): 320-329.

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Cloning and Sequence Analysis of MC4R Gene in Luchuan Pig

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