鸡类KISS-1基因克隆及EST序列表达验证分析  

Cloning of the Chicken KISS-1 Like Gene and the Expression Analysis of EST Sequence

作　　者：韩威[1,2] 苏一军[1] 朱云芬[1] 宋迟[1] 李国辉[1] 屠云洁[1] 张会永[1] 殷建梅[1] 

机构地区：[1]中国农业科学院家禽研究所,江苏扬州225125 [2]南京农业大学,江苏南京210095

出　　处：《中国家禽》2014年第16期12-16,共5页China Poultry

基　　金：国家自然科学基金项目(31201799)

摘　　要：性成熟启动是一个复杂的生理过程,受＂下丘脑-垂体-性腺＂轴控制。决定性成熟启动的关键性事件是下丘脑促性腺激素释放激素（GnRH）脉冲式释放增加,KISS-1基因编码产物Kisspeptin通过其受体GPR54能够直接刺激GnRH释放,进而启动性成熟进程,KISS-1/GPR54系统被认为是性成熟启动的关键看门基因。禽类的KISS-1基因在GenBank中尚未被清晰注释。本文以文昌鸡为素材,采用比较基因组学方法分析了不同物种KISS-1及其上、下游基因在染色体上的分布,确定了鸡chr26：1522493-1532710区域为候选片段,通过PCR扩增和克隆测序获得了该候选片段完整序列（命名为类KISS-1序列）;同时,采用BLAST程序搜索出9个同源性在50%～85%的EST序列,通过拼接得到3个延伸EST序列。表达验证分析表明,3个延伸EST序列在文昌鸡下丘脑组织cDNA文库中均有表达。Puberty onset of animals is a complex biological process and under the control of the hypothatamicpitutay-gonadal axis. The increased pulsatile release of gonadotrophin releasing hormone（GnRH）from the hypothalamus is the determined event. Kisspeptins,encoded by KISS-1 gene,can directly promote the release of GnRH through GPR54 receptors and start the puberty processing. So,KISS-1 and GPR54 system is treated as the gatekeeper of puberty onset. However,the avian KISS-1 sequences are not interpreted clearly in GenBank database. In this study,Wenchang chicken was used as the material,and the chr26：1522493-1532710 region was determined as the candidated fragments by analyzing the distribution of KISS-1 and its near genes on the chromosome in different species. The intact sequence was acquired by PCR and sequencing,called KISS-1 like sequence.Nine ESTs with 50% to 80% similarity to the candidated fragments were found,and three prolonged ESTs were acquired by split joint. The prolonged ESTs all expressed in the hypothatamic cDNA bank of Wenchang chicken.

关 键 词：性成熟启动 鸡 KISS-1基因 EST序列 

分 类 号：Q78[生物学—分子生物学]