牛卵泡抑素基因克隆及真核表达载体构建  

Gene Cloning and Construction of Bovine Follistatin Expression Vector

作　　者：康峰[1] 苏广华[1] 苏建国[2] 段彪[1] 王子东[1] 李光鹏[1] 

机构地区：[1]内蒙古大学生殖生物学及生物技术教育部重点实验室,内蒙古呼和浩特010021 [2]西北农林科技大学动物科技学院,陕西杨凌712100

出　　处：《中国牛业科学》2011年第5期9-12,共4页China Cattle Science

基　　金：高产转基因肉牛新品种培育(编号:2008ZX08007-002)

摘　　要：[目的]克隆牛的卵泡抑素基因(Follistatin,FSTN)基因,构建真核表达载体。[方法]用Trizol法从牛的卵巢中提取总RNA,反转录成cDNA,用带有酶切位点牛FSTN的特异性引物扩增其完整编码区序列,连接到T载体、测序,序列无误后亚克隆入真核表达载体pIRES2-AcGFP1中,酶切及PCR鉴定载体。[结果]经酶切及PCR鉴定表明成功构建了真核表达载体pIRES2-AcGFP1-FSTN。[结论]成功构建了真核表达载体pIRES2-AcGFP1-FSTN,为促进肌肉生长的转基因优质肉牛品种培育奠定了基础。【Objective】To clone and construct the eukaryotic expression vector containing the bovine follistatin cDNA sequence.【Methods】Total RNA was extracted from bovine ovary by Trizol total RNA Extract Kit,and the whole coding region of bovine follistatin cDNA gene was cloned by RT-PCR using specific primers containing restriction enzyme cutting site.The purified bovine follistatin cDNA was inserted into pMD18-T vector and was then sequenced,then subclone the correct follistatin cDNA to eukaryotic expression vector pIRES-AcGFP.Double-enzyme cleavage and PCR test were used to identify the vector.【Results】Double-enzyme cleavage and PCR test showed that FSTN was successfully cloned into eukaryotic expression vector.【Conclusion】We successfully constructed the eukaryotic expression vector pIRES2-AcGFP1-FSTN.This study provided the foundation for fostering high beef quality transgenic cattle of promoting muscle growth.

关 键 词：牛卵泡抑素基因 基因克隆 真核表达载体构建 

分 类 号：S823.2[农业科学—畜牧学]