大鼠神经球蛋白基因的克隆及其真核表达载体构建  

Cloning of rat neuroglobulin gene and construction of its eukaryotic expression vector

作　　者：朱继[1] 唐文渊[1] 

机构地区：[1]重庆医科大学附属第一医院神经外科,重庆400016

出　　处：《第三军医大学学报》2007年第2期144-146,共3页Journal of Third Military Medical University

基　　金：重庆市卫生局资助项目(06-2-177)~~

摘　　要：目的克隆Wistar大鼠神经球蛋白(neuroglobulin,NGB)基因并构建其真核表达载体,为进一步研究其功能奠定基础。方法从雄性Wistar大鼠脑组织中提取总RNA,经逆转录PCR得到cDNA,测序并利用BLAST工具对比证明该基因序列正确后,亚克隆入pCDNA3.1(+)载体中,并通过双酶切和测序确定序列和阅读框正确。结果克隆了Wistar大鼠的神经球蛋白基因,有4个碱基发生了突变;成功构建了真核表达载体。结论Wistar大鼠脑组织中有NGB表达,成功克隆了该基因并构建了其真核表达载体。Objective To clone vector. Methods The total RNA was rat neuroglobulin （NGB） gene and construct its eukaryotic expression extracted from Wistar rat brain and the full length cDNA encoding NGB was obtained by RT-PCR. After the sequence was confirmed by sequencing and BLAST, it was inserted in the eukaryotic expression vector pCDNA3.1 （ ＋ ）, then the sequence and reading frame were confirmed by two restriction endonucleases and sequencing. Results The NGB gene was cloned with four bases mutated and its eukaryotic expression vector was constructed. Conclusion NGB expressed in Wistar rat brain. NGB gene was successfully cloned and inserted in eukaryotic expression vector.

关 键 词：神经球蛋白基因 脑组织 克隆 表达载体 

分 类 号：Q512.2[生物学—生物化学] Q782