人脑源性神经营养素-6基因的克隆及在原核细胞中的表达  

Cloning of human brain-derived neurotrophin-6 gene and its expression in procaryotic cell

作　　者：张承武[1] 蔡青松[2] 庄旭 翟朝阳 郑煜 

机构地区：[1]四川大学华西基础医学与法医学院生理学教研室,成都610041 [2]四川大学华西基础医学与法医学院生物化学与分子生物学教研室,成都610041

出　　处：《中华医学遗传学杂志》2002年第6期476-478,共3页Chinese Journal of Medical Genetics

基　　金：四川大学华西基础医学与法医学院生物化学与分子生物学开放实验室客座研究基金~~

摘　　要：目的　克隆人脑源性神经营养素 - 6 (neurotrophin- 6 ,NT- 6 )基因 ,并观察其在大肠杆菌中的表达情况。方法　从流产胎儿大脑皮层提取总 RNA,用逆转录聚合酶链反应扩增得到人的 NT- 6基因c DNA片段 ;经酶切回收后克隆进 p BK- CMV质粒 ,构建 NT- 6的表达载体。通过诱导 ,使 NT- 6基因在大肠杆菌中表达。结果　分离克隆了人脑源性 NT- 6基因 ,含该基因的大肠杆菌在异丙基 -β- D-硫代半乳糖苷诱导的情况下 ,表达了特异性的蛋白质。结论　人脑源性 NT- 6基因的克隆表达为进一步研究 NT- 6的结构、功能及临床应用奠定了基础。Objective To clone human brain-derived neurotrophin-6(NT-6) gene and to observe its expression in the procaryotic cell. Methods Total RNA was extracted from aborted antenatal cerebral cortex, and cDNA fragment of NT-6 was amplified through reverse transcript-polymerase chian reaction. After being incised and recovered, the NT-6 gene was cloned into pBK-CMV plasmid to construct a NT-6 gene expression vector. Expression of NT-6 gene in Escherichia coli was studied after being induced by isopropyl β-D-thiogalactoside(IPTG). Results The NT-6 gene expression vector was constructed and Escherichia coli with recombinant vector expressed specific protein after induction by IPTG. Conclusion The cloning of human brain-derived NT-6 gene provides a basis for further studying the structure, function and clinical application of NT-6.

关 键 词：人神经营养素-6 克隆 原核表达 蛋白质 

分 类 号：R346[医药卫生—基础医学]