庞实锋,付宏岐,郑克勤,曹定国,周汝滨
PANG Shifeng1,FU Hongqi2,ZHENG Keqin1,CAO Dingguo1,ZHOU Rubin1 (1.Department of Biology

• 1:广东医学院生物学教研室
• 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

摘要(Abstract)：

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(KeyWords)： 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
human acidic fibroblast growth factor;gene cloning;temperature-dependent expression;polymerase chain reaction

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 庞实锋,付宏岐,郑克勤,曹定国,周汝滨
PANG Shifeng1,FU Hongqi2,ZHENG Keqin1,CAO Dingguo1,ZHOU Rubin1 (1.Department of Biology

DOI: 10.19367/j.cnki.1000-2707.2011.04.007

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