睫状神经营养因子的cDNA克隆、表达、纯化及其生物活性鉴定  

Cloning expression and purification of human Ciliary neurotrophic factor cDNA and its function identification

作　　者：江国春[1] 袁丽珍[1] 贾向旭[1] 

机构地区：[1]军事医学科学院放射医学研究所,北京100850

出　　处：《基础医学与临床》2000年第1期27-30,共4页Basic and Clinical Medicine

摘　　要：反转录ＰＣＲ扩增人睫状神经营养因子（ｈｕｍａｎｃｉｌｉａｒｙｎｅｕｒｏｔｒｏＰｈｉｃｆａｃｔｏｒ，ｈＣＮＴＦ）ｃＤＮＡ后，克隆进ｐＢＶ２２０，在大肠杆菌中实现原核高效表达，ＳＤＳ－ＰＡＧＥ观察到分子量约为２４ｋＤ的预期表达产物，薄层扫描确定其表达量为２６％，制备电泳和凝胶过滤纯化后纯度达到９８％，生物活件分析表明，表达产物能刺激１０日龄鸡胚背根神经节细胞的生长，促进ＣＦＵ－ＧＭ的增殖。After obtaining the optimum expression conditions, the cDNA of human ciliary neurotrophic factor (hCNTF) was cloned, and highly expressed in E.Coli. Total RCA was extracted from human fetus sciatic nerve tissues, and the CNTF cDNA was obtained by RT-PCR. Then the CNTF cDNA was cloned into PUC 18 and PBV220 respectively. After highly expressed in E Colt., rhCNTF was purified with preperative electrophoresis and gel filtration, which can survive the growth of E10 DRG neurons and CFU-GM.

关 键 词：睫状神经营养因子 CNTF 基因克隆 CDNA PCR 

分 类 号：Q523[生物学—生物化学] Q593.2