人胰岛素样生长因子-1基因的克隆、表达及生物活性分析  

Clonging, Expression and Biological Activity of Human Insulin-like Growth Factor-1

作　　者：祁雅慧[1] 王雅梅[1] 孙丽翠[1] 司杨[1] 闫豫东[1] 张静宜[1] 邴国英 

机构地区：[1]首都医科大学实验中心 [2]美国Kentucky大学解剖和神经生物学部

出　　处：《首都医科大学学报》2004年第2期154-158,共5页Journal of Capital Medical University

基　　金：北京市科学技术委员会基金 ( 95 40 2 480 0 )资助项目

摘　　要：采用RT PCR方法 ,从人扁桃体组织中扩增得到人胰岛素样生长因子 1 (IGF 1 )cDNA ,利用基因重组技术将该基因片段重组于pcDNA3 .1 ( +)真核表达载体上 ,成功构建了 pcDNA/I重组表达载体。应用脂质体介导的基因转移技术将重组质粒体外转染至人脐静脉内皮细胞 (HUVEC)。MTT法检测结果表明 :重组质粒转染能明显促进内皮细胞的分裂增生。将重组质粒通过脂质体介导 ,体外转染至人肾细胞 2 93细胞系进行表达 ,经G41 8筛选获得稳定表达的重组质粒细胞克隆 ,表达产物经鸡胚绒毛尿囊膜试验表明有促血管生成活性。此结果为研究IGF 1对血管内皮细胞作用的分子机制 ,以及利用IGF 1基因治疗肢体及冠状动脉缺血性疾病等的研究奠定了实验基础。The human insulin-like growth factor-1(IGF-1) cDNA was amplified by RT-PCR from human tonsil tissue. Sequenced, the IGF-1 cDNA was inserted into eukaryotic expression vector pcDNA3.1(+). The recombinant plasmid pcDNA/I was transferred into HUVEC cells mediated by liposome. The activity of IGF-1 was detected by MTT. The results showed that IGF-1 protein was expressed in HUVEC cells 72 h after gene transference and it had good biological activity to stimulate HUVEC proliferation. pcDNA/I was transferred into 293 cells mediated by liposome. Screened with G418, we got positive cell which can express recombinant protein of IGF-1 stably. Chick charioallantoic membrane(CAM) bioassay show that the recombinant protein has biological activity to promote blood vessel formation.

关 键 词：人胰岛素样生长因子-1基因 克隆 表达 生物活性 

分 类 号：Q785[生物学—分子生物学]