山羊MSH4和MSH5基因的cDNA克隆、序列分析及组织表达

doi:10.13560/j.cnki.biotech.bull.1985.2016.03.016

摘要/Abstract

摘要： 以低繁藏山羊和高繁金堂黑山羊为研究对象, 分别提取处于发情期的5只藏山羊和5只金堂黑山羊的卵巢、子宫、输卵管、垂体的总RNA, 并通过RT-PCR技术对MSH4、MSH5基因cDNA进行克隆、序列分析, 以Real-time PCR技术对其进行组织表达研究。结果表明, 藏山羊和金堂黑山羊MSH4基因编码区均长2 499 bp, 编码832个氨基酸, 两品种基因编码区有5处碱基不同, 并导致3处氨基酸的差异；MSH5基因编码区均长2 496 bp, 编码831个氨基酸, 两品种基因编码区有9处碱基不同, 并导致5处氨基酸的差异。藏山羊MSH4基因编码区核苷酸序列与金堂黑山羊、山羊、绵羊、牛、马、小鼠、褐家鼠、人的同源性分别为：99.8%、99.8%、99.4%、98.1%、94.4%、85.1%、84.7%和93.5%；藏山羊MSH5基因编码区核苷酸序列与金堂黑山羊、山羊、牛、家犬、小鼠、褐家鼠、人的同源性分别为：99.6%、99.6%、97.3%、88.0%、85.8%、85.3%和90.2%。MSH4和MSH5基因mRNA在两个山羊品种的卵巢、子宫、输卵管、垂体中均有表达, 但两品种间无显著性差异(P>0.05)。说明MSH4和MSH5基因在动物进化中比较保守, 与山羊多羔性状的相关性有待进一步研究。

关键词: 藏山羊, 金堂黑山羊, Msh4, Msh5, 克隆, 定量PCR

Abstract: The total RNA of ovary, endometrium, oviduct and pituitary extracted from 5 Tibetan goats and 5 Jintang black goats in the period of estrus were used to clone and analyze cDNA sequences of gene MSH4 and MSH5 by RT-PCR, and their expression levels in different tissues were determined by real-time PCR. The result showed that the coding region of gene MSH4 from Tibetan goat and Jintang black goat were 2 499 bp, encoding 832 amino acids；there were 5 base variations between two breeds, leading to 3 differences of amino acid. The coding region of gene MSH5 from Tibetan goat and Jintang black goat were 2 496 bp, encoding 831 amino acids；there were 9 base variations between two breeds, leading to 5 differences in amino acid. The nucleotide sequences of coding region of gene MSH4 from Tibetan goat were 99.8%, 99.8%, 99.4%, 98.1%, 94.4%, 85.1%, 84.7% and 93.5% homology with that of Jintang black goat, Capra hircus, Ovis aries, Bos taurus, Equus caballus, Mus musculus, Rattus norvegicus, and Homo sapiesn, respectively. The nucleotide sequences of coding region of gene MSH5 from Tibetan goat were 99.6%, 99.6%, 97.3%, 88.0%, 85.8%, 85.3% and 90.2% homology with that of Jintang black goat, C. hircus, B. taurus, Canis lupus familiaris, M. musculus, R. norvegicus, and H. sapiens, respectively. The mRNA of gene MSH4 and MSH5 were all expressed in ovary, endometrium, oviduct and pituitary of two breeds of goat, but there was no significant difference between two breeds(P>0.05). The results reveal that both gene MSH4 and MSH5 are conservative in the course of animal evolution, and the role of their effect on prolificacy of goats needs to be further studied.

Key words: Tibetan goat, Jintang black goat, MSH4, MSH5, cloning, qPCR

郑杰,刘霜,罗斌,胡亮,杨珂伟,字向东. 山羊MSH4和MSH5基因的cDNA克隆、序列分析及组织表达[J]. 生物技术通报, 2016, 32(3): 98-104.

ZHENG Jie, LIU Shuang, LUO Bin, HU Liang, YANG Ke-wei, ZI Xiang-dong. cDNA Cloning, Sequence Analysis and Tissue Expression of Gene MSH4 and MSH5 in Goats[J]. Biotechnology Bulletin, 2016, 32(3): 98-104.

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