山羊INHA和INHBA基因的cDNA克隆、序列分析及组织表达研究

cDNAcloning,sequence analysis and tissue expression of INHA and INHBA genes in goats

分别提取处于同一发情周期的5只低繁藏山羊和5只高繁金堂黑山羊的卵巢、垂体的总RNA,并通过RT-PCR技术对INHA、INHBA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INHA基因编码区均长1083bp,编码360个氨基酸,两品种基因编码区有7处碱基不同,并导致3处氨基酸的差异;INHBA基因编码区均长1278bp,编码425个氨基酸,两品种基因编码区有4处碱基不同,并导致1处氨基酸的差异.藏山羊INHA基因编码区核苷酸序列与金堂黑山羊、绵羊、牛、野猪、小鼠、褐家鼠、人的同源性分别为:99.4%、98.9%、95.8%、88.6%、81.0%、79.5%和84.8%;藏山羊INHBA基因编码区核苷酸序列与金堂黑山羊、绵羊、牛、野猪、小鼠、褐家鼠、人的同源性分别为:99.7%、99.4%、98.1%、91.7%、88.0%、88.5%和91.2%.INHA和INHBA基因mRNA在两个山羊品种的卵巢、垂体中均有表达,但两品种间无显著性差异(P>0.05).说明INHA和INHBA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.

Ovary and pituitary samples were collected from five non-prolific Tibetan goats and five prolific Jintang black goats in estrus,to clone and analyze sequences of INHA and INHBA cDNA by RT-PCR,and their mRNA expression levels were determined by real-time PCR. The result showed that the coding region of Tibetan goat and Jintang black goat INHA gene were1083 bp long,encoding 360 amino acids. There were seven base changes between the two breeds,leading to three differences in amino acid. The coding region of Tibetan goat and Jintang black goat INHBA gene were 1278 bp long,encoding 425 amino acids. There were four base changes between the two breeds,leading to one amino acid difference. The homologies of coding region of INHA gene between Tibetan goat,Jintang black goat,Ovis aries,Bos taurus,Sus scrofa,Mus musculus,Rattus norvegicus,Homo sapiens were 99. 4%,98. 9%,95. 8%,88. 6%,81. 0%,79. 5% and 84. 8%,respectively. The homologies of coding region of INHBA gene between Tibetan goat,Jintang black goat,Ovis aries,Bos taurus,Sus scrofa,Mus musculus,Rattus norvegicus,Homo sapiens were 99. 7%,99. 4%,98. 1%,91. 7%,88. 0%,88. 5% and 91. 2%,respectively. The INHA and INHBA mRNA were expressed in ovary and pituitary in both goat breeds,but there was no significant difference between the two breeds（ P 〉0. 05）. The result showed that both INHA and INHBA genes were conservative in animal evolution,and their effect on prolificacy in goats needs to be further studied.