人神经营养因子-3基因在大肠杆菌中的克隆和表达  

Gene Cloning and Expression of Human Neurotrophin-3 in E.Coli

作　　者：章建程[1] 殷明[1] 戴建凉[1] 林卿[1] 余浩[1] 刘忠权[1] 

机构地区：[1]海军医学研究所,上海市200433

出　　处：《海军医学杂志》1999年第2期125-127,共3页Journal of Navy Medicine

摘　　要：目的 :获得人神经营养因子 - 3 (hNT - 3 )在大肠杆菌中的克隆并表达。方法 :采用PCR方法从 2 0周人胎脑cDNA文库中扩增hNT - 3基因 ,通过双酶切法在M 13mp18载体上克隆获得全长基因 ,将正确全长基因插入PBV2 2 0载体 ,通过选择合适宿主菌获得hNT - 3的高效表达。结果 :获得全长基因完全正确的hNT - 3基因 ,获得占细菌总蛋白 3 0 %的hNT - 3表达菌株。结论 :本研究获得了序列正确的hNT - 3基因克隆 ,并实现了在大肠杆菌中的高效表达。Objective: To obtain hNT-3 gene and express it in E.coli. Methods: The human fetal brain cDNA library was chosen as the template to obtain hNT-3 gene through PCR. The PCR products, which had been cut by a suitable enzyme, were cloned into M13mp18 vector. It was then cloned into expression vector PBV220. The rhNT-3 was highly expressed in E. coli. Results: The correct full-length hNT-3 gene was obtained, and the expressed hNT-3 amounts to 30% of the total bacterial protein. Conclusion: A confirmed hNT-3 clone was obtained and highly expressed in E. coli.

关 键 词：人神经营养因子-3基因 PCR扩增 基因重组 基因表达 

分 类 号：Q785[生物学—分子生物学]