摘要
目的构建人类成熟型TGF-β1(hTGF-β1)基因的表达载体,探索TGF-β1在大肠埃希菌中的表达,获得重组表达载体pET32a-hTGFβ1。方法Trizol法抽提人新鲜胎盘组织的总RNA,RT-PCR法扩增人成熟型TGF-β1基因,PCR产物纯化后经BglⅡ和HindⅢ双酶切后与相同酶切的pET32a(+)载体连接,构建重组载体pET32a-hTGF-β1。结果成功构建了hTGF-β1重组表达载体pET32a-hTGFβ。结论hTGF-β1基因的原核表达载体的成功构建,为下一步表达、纯化该基因的蛋白产物以及hTGF-β1多克隆抗体的制备提供了依据。
Objective To construct expression vector of human maturate TGF-β1（hTGF-β1） in E. coll. obtaining recombinant expression vector pET32a-hTGFβ1. Methods The human mature TGF-β1 was amplified by RT-PCR from the total RNA of healthy person＇s placenta by trizol agent, then both of the prokaryotic expression vector pET32a（ ＋ ） and hTGF-β1 gene were digested with Bgl Ⅱ and Hind Ⅲ. After purification, hTGF-β1 gene was inserted into pET32a（ ＋ ）, construction recombinant vector pET32a-hTGF-β1. Results Succeess to construct recombination expression vector pET32a-hTGFβ1. Conclusion Our successful cloning lay a basis for expression and purification of hTGF-β1 protein and the production of hTGF-131 polyelonal antibody.
出处
《中国现代药物应用》
2008年第24期8-9,共2页
Chinese Journal of Modern Drug Application
