胶质细胞源性神经营养因子基因重组腺病毒穿梭质粒的构建及鉴定  

Construction and identification of recombinant shuttle adenovirus plasmid containing glialcellline derived neurotrophic factor.

作　　者：孙念峰[1] 舒晓刚[1] 张景辉[1] 王国斌[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院普外科,武汉430022

出　　处：《中国综合临床》2006年第4期351-353,共3页Clinical Medicine of China

基　　金：国家自然科学基金(30371397)

摘　　要：目的克隆大鼠胶质细胞源性神经营养因子(GDNF)基因,构建含有GDNF基因的重组腺病毒载体,为转染神经干细胞和基因治疗奠定基础。方法从大鼠脑组织中提取总RNA,用逆转录聚合酶链反应方法扩增出约660bp的片段,并将该片段克隆到载体pAdTrack-CMV中,进行酶切鉴定和序列分析。结果证实成功构建了含有GDNF基因的重组腺病毒质粒。结论克隆到正确的大鼠GDNF基因,并构建出重组质粒pAd-GDNF,为下一步重组腺病毒颗粒和基因治疗打下基础。Objective To clone rat glialeellline derived neutrophie factor gone （GDNF） and construct recombinant adenovirus vector containing GDNF gent, which was then transfected into neural stem cells. Methods talRNA was extracted from rat brain tissue. A660bp positive eDNA was obtained by RT-PCR and then was cloned into the shuttle plasmid pAdTract-CMV. The cloned fragment, was sequenced and identified by restriction enzyme. Results Recombinant adenovirus plasmid containing GDFN gene was obtained successfully. Conclusion A recombinant adcnovirus plasmid containing GDNF is coustruetcd suecessfuUy which lays the foundation for further study on aene theraDv and recombinant adenovirus olasmid.

关 键 词：基因克隆 胶质细胞源性营养因子 逆转录聚合酶链反应 腺病毒载体 序列分析 

分 类 号：R346[医药卫生—基础医学]