Prepro-neurturin基因的克隆及其在COS-7中的表达  

Cloning of prepro-neurturin gene and its expression in COS-7 cells

作　　者：孙秀[1] 糜军[2] 陈生弟[1] 陈诗书[2] 

机构地区：[1]上海第二医科大学附属瑞金医院神经科 [2]上海第二医科大学分子生物学实验室,上海200025

出　　处：《中国神经科学杂志》2001年第4期358-361,共4页

基　　金：国家重点基础研究规划"脑功能和脑重大疾病的基础研究"(G19990 5 40 0 8);国家自然科学基金 ( 39970 2 6 3);上海市卫生系统百名跨世纪优秀学科带头人培养计划 ( 97BR0 0 1);上海市科委"启明星后"计划 ( 97QMB14 10 )资助项目

摘　　要：Neurturin (NTN)是对多巴胺能神经元有营养、支持和保护作用的神经营养因子 ,是帕金森病治疗的潜在有效因子。本研究用RT PCR方法获取prepro neurturincDNA ,并克隆至pBluescriptⅡSK(+)载体中 ,经酶切及测序鉴定正确 ;再将prepro neurturin基因克隆至 pcDNA3真核表达载体中 ,经Lipofectamine转染 ,在COS 7细胞中瞬间表达 ,经NorthernBlot和WesternBlot印迹分析 ,鉴定其mRNA和蛋白表达正确。用表达NTN的条件培养基处理原代培养胚胎中脑神经元 ,发现其有促进神经元存活和发育功能。Neurturin(NTN), a recently found neurotrophic factor, exhibits survival promoting and protective actions on dopaminergic neurons, thus making NTN a potential therapeutic factor for Parkinson's disease. In the present study, we cloned the mouse prepro neurturin by RT PCR, subcloned it into pBluescriptⅡ SK(+), and identified the right clone by digestion of restriction enzyme and sequencing. Then we transfected pcDNA3/prepro neurturin into COS 7 cells with lipofectamine, and identified the mRNA and protein expression of prepro NTN gene by Northern Blot and Western Blot respectively.It was found that NTN can promote survival and growth of neurons when culturing primary embryonic mesencephalic neurons with conditioned medium contained NTN.

关 键 词：Prepro-neurturin 多巴胺能神经元 帕金森病 COS-7 基因克隆 

分 类 号：R346[医药卫生—基础医学] R742.5