实时荧光定量PCR检测鸡生长激素基因方法的建立  

Development of a Real-time FQ-PCR Method for Detection of Growth Hormone Gene in Chickens

作　　者：崔帅[1] 李娅[1] 金元昌[2] 韦平[1] 黄亮[1] 

机构地区：[1]广西大学养禽与禽病研究所,广西南宁5300042 [2]湖南科技大学生命科学学院,湖南湘潭411201

出　　处：《中国家禽》2012年第11期36-39,共4页China Poultry

基　　金：广西科技攻关项目(桂科攻10100005-10);国家自然科学基金(30460099)

摘　　要：本研究旨在建立定量分析鸡生长激素(Growth hormone,GH)基因mRNA表达水平的实时荧光定量PCR方法,为鸡GH基因mRNA表达水平的定量分析及探讨其与马立克氏病遗传抗性间关系奠定基础。根据GenBank数据库中鸡的GH基因序列,使用Oligo7软件在该基因保守区域设计合成一对引物并扩增GH基因片段,经回收纯化后连接到pGM-T载体,成功构建了含有GH基因片段的pGM-T-GH标准质粒。在此基础上,通过SYBR GreenⅠ染料法建立了pGM-T-GH重组质粒的实时荧光定量PCR标准曲线及其回归方程,成功建立了实时荧光定量PCR检测鸡GH基因的方法。This study was aimed to establish a real-time FQ-PCR method for analysis of growth hormone （GH） gene′s mRNA expression , while laid the foundation for quantitative analysis of GH gene′s mRNA expression and the relationship with genetic resistance of Marek′s disease. According to the published sequences of chicken GH gene in GenBank , a pair of primers was designed by Oligo 7 according to the conserved region and amplified specific fragment of GH gene. Target fragment was linked to the pGM-T vector after purification , and then the standard sample of plasmid pGM-T-GH was constructed successfully.Standard curve and regression equation of pGM-T-GH in real-time FQ-PCR based on SYBR GreenⅠ method was established , and successfully developed a real-time FQ-PCR method for the detection of GH gene chickens.

关 键 词：GH基因 实时荧光定量PCR 重组质粒 标准曲线 

分 类 号：S513[农业科学—作物学]