haFGF改构体的温控表达及其活性测定  

Temperature-regulated Expression of Human Acidic Fibroblast Growth Factor Remodeling Gene and Detection of Its Activity

作　　者：庞实锋[1] 付宏岐[2] 郑克勤[1] 曹定国[1] 周汝滨[1] 

机构地区：[1]广东医学院生物学教研室,广东湛江524023 [2]吉林农业大学生物反应器与药物开发教育部工程研究中心,吉林长春130118

出　　处：《贵阳医学院学报》2011年第4期351-354,358,共5页Journal of Guiyang Medical College

摘　　要：目的:构建温控表达载体,采用温控表达haFGF,为降低其生产成本打下基础。方法:采用PCR法扩增截去其N端19个氨基酸的haFGF DNA,克隆到温控表达载体PBV220中,然后转化进BL21(DE3)Star plysS中进行温控表达。结果:经SDS-PAGE和Western blot分析表明,温控表达载体表达的haFGF具有其原来的免疫原性,MTT活性检测结果表明,haFGF纯化产物与野生型haFGF活性相当,差异无显著性(P>0.05)。结论:成功构建了haFGF的温控表达载体,为提高目的蛋白的表达量和降低其生产成本打下基础。Objective：To construct temperature-regulated expression vector,and to express human acidic fibroblast growth factor（haFGF） in a temperature-regulated manner,and so as to lay a foundation for cutting down cost of haFGF production.Methods：HaFGF truncated by 19 amino acids in N-terminal was amplified with polymerase chain reaction（PCR）,cloned into temperature-regulated expression vector PBV220,and then transformed into E.coli.BL21（DE3）Star plysS to express under induction of temperature.Results：SDS-PAGE and Western blot analysis showed that haFGF-19 had its original immunogenicity.MTT results indicated that there was no significant difference of activities between haFGF-19 and wild-type aFGF.Conclusions：Temperature-regulated expression vector is successfully constructed.This study lays a foundation for enhancing expression level of haFGF and cutting down cost of production.

关 键 词：酸性成纤维细胞生长因子 基因克隆 温控表达 聚含酶链反应 

分 类 号：Q786[生物学—分子生物学]