山羊促性腺激素释放激素1基因外显子克隆及序列分析  

Cloning and Sequence Analysis on Exons of Gonadotropin Releasing Hormone 1 Gene in Goats

作　　者：闫艳[1] 储明星[2] 狄冉[2] 方丽[2] 王凭青[1] 

机构地区：[1]重庆大学生物工程学院,重庆400030 [2]中国农业科学院北京畜牧兽医研究所,农业部畜禽遗传资源与利用重点开放实验室,北京100193

出　　处：《中国畜牧兽医》2009年第10期75-79,共5页China Animal Husbandry & Veterinary Medicine

基　　金：国家高技术研究发展计划(863)(2006AA10Z139);国家现代农业产业技术体系建设专项资金(农科教发[2008]10号);国家重点基础研究发展计划(973)(2006CB102105);国家自然科学基金项目(30871773);北京市自然科学基金项目(6062023);中国农业科学院首批优秀科技创新团队专项资助

摘　　要：促性腺激素释放激素1（gonadotropin-releasing hormone 1,GnRH1）是GnRH存在于脑下垂体的主要形式,它在调节生殖内分泌系统产生促性腺激素方面起着主要作用。根据牛GnRH1基因全序列设计3对引物,采用PCR-SSCP技术检测GnRH1基因外显子2、3和4在高繁殖力山羊品种（济宁青山羊）、中等繁殖力山羊品种（波尔山羊）和低繁殖力山羊品种（内蒙古绒山羊和安哥拉山羊）中的单核苷酸多态性,分析该基因对山羊高繁殖力的影响;并对山羊GnRH1基因3个外显子进行克隆测序,推导出编码的氨基酸序列,然后将山羊核苷酸和氨基酸序列与牛、人、狗、黑猩猩、猴、大鼠和小鼠7个物种的序列进行同源性比较。结果表明,在4个山羊品种中均未检测到GnRH1基因3个外显子的多态性;8个物种的核苷酸和氨基酸序列的同源性分别为73.6%~99.3%和62.0%~97.8%。可见,哺乳动物GnRH1基因外显子2、3和4序列保守性较强,该区域可能不是影响山羊高繁殖力的功能结构域。As gonadotropin releasing hormone 1 （GnRH1） is the main form of GnRH present in the pituitary, it seems ap parent that it is responsible for regulating the reproductive endocrine system to stimulate gonadotropins. The GnRH1 gene was studied as a candidate gene for high prolificacy in some goat breeds （Capra hircus）. Single nucleotide polymorphisms （SNPs） of three exons （exons 2, 3 and 4） of GnRH1 gene were detected in high prolificacy breed （Jining Grey goat）, moderate prolifi cacy breed （Boer goat） and low prolificacy breeds （Inner Mongolia Cashmere and Angora goats） by polymerase chain reaction- single strand conformation polymorphism （PCR SSCP）. No polymorphism was detected in the amplified region in the four goat breeds tested. DNA fragments of exons 2, 3 and 4 of Jining Grey goat GnRH1 gene were cloned into pSRUE T vector. The positive clones were further identified by PCR. The nucleotide sequences were detected and the amino acid sequences of these three fragments were deduced. The comparison of the fragments of all three coding regions of Jining Grey goat GnRH1 gene with those of cattle, human, dog, chimpanzee, monkey, rat and mouse GnRH1 gene showed that the homologies of the nucleo tide sequences and amino acid sequences of the fragments among these eight species were from 73. 6% to 99.3% and from 62.0% to 97.8% respectively. The results indicated that the exons 2, 3 and 4 of GnRH1 gene were highly conserved among mammals, and may not be the functional domains that affect the high prolificacy in goats.

关 键 词：山羊 促性腺激素释放激素1基因 PCR-SSCP 

分 类 号：Q785[生物学—分子生物学]