人睫状神经营养因子的克隆与原核表达  

Cloning and expression of human ciliary neurotrophic factor cDNA from fetus

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

出　　处：《军事医学科学院院刊》1998年第2期118-121,124,共5页Bulletin of the Academy of Military Medical Sciences

摘　　要：目的：克隆人睫状神经营养因子基因ｃＤＮＡ，并探索其在大肠杆菌中高效表达。方法：提取总ＲＮＡ，逆转录聚合酶链反应（ＲＴ－ＰＣＲ），序列测定正确后原核表达并用制备电泳纯化。结果：克隆了人睫状神经营养因子基因ｃＤＮＡ，实现了其在大肠杆菌中的高效表达，表达量至少达到２６％，制备电泳纯化后，重组蛋白纯度达到９５％。结论：从胎儿坐骨神经组织中克隆到人睫状神经营养因子基因ｃＤＮＡ。Objective: To clone the cDNA of human ciliary neurotrophic factor (hCNTF) and to express it in E.coli. Methods: Total RNA was extracted from cells and tissues and, after RT PCR and sequence analysis, expressed in prokaryotes and purified . Results: cDNA of CNTF was cloned and highly expressed in E.coli, with an expression level of 26%. After purification with preparative electrophoresis, the purity of the recombinant protein was 95%. Conclusion: The cDNA of CNTF was cloned from fetal sciatic nerve tissues and highly purified recombinant protein was obtained after purification with preparative electrophoresis.

关 键 词：神经生长因子 聚合酶链反应 基因表达 克隆 CNTF 

分 类 号：Q78[生物学—分子生物学]