生命科学研究2011，Vol.15Issue(5)：438-442,5.
GOLPH2的原核表达、纯化、抗体制备和初步应用
Expression, Purification, Antibody Preparation and Preliminary Application of Tumor Protein GOLPH2
摘要
Abstract
Recombinant expression of G0LPH2 and preparation of anti-G0LPH2 polyclonal antibody that may be a serologic marker of hepatocellular carcinoma(HCC) and clinical significance of the feasibility of the basis, golphl was amplified from HepG2 cells and cloned into the prokaryotic expression vector pET21a(+)-TRX, then expressed in E. Coli BL21 (DE3) which was induced by isopropylthio-jS-D-galactoside (IPTG). The recombinant protein G0LPH2 was purified by His-tag magnetic bead purification kit and detected by SDS-PAGE. Polyclonal antibodies was developed by immunizing BALB/c mice with purified recombinant protein, The specificity and titer of the antibody in anti-sera were determined by Western blot and ELJSA respectively. The prokaryotic expression plasmid pET21a (+)-TRX-G0LPH2 was successfully constructed. The recombinant protein TRX-G0LPH2 could be expressed in abundance in the soluble form and got the purified purpose protein of 52 kD. SDS-PAGE and Western blot analysis showed that GOLPH2 protein was successfully expressed in BL21 (DE3). It was showed that antiserum can specifically identify the 52 kD recombinant protein, 73 kD cell lysates and serum protein specifically by Western blot analysis. The polyclonal antibody a-gainst G0LPH2 protein was successfully prepared, and can be used for follow-up examination.关键词
肝细胞癌/蛋白质表达/GOLPH2/多克隆抗体Key words
hepatocellular carcinoma/ expression of protein/ G0LPH2/ polyclonal antibody分类
医药卫生引用本文复制引用
李峰,刘艳红,谢平丽,李跃辉,李官成..GOLPH2的原核表达、纯化、抗体制备和初步应用[J].生命科学研究,2011,15(5):438-442,5.基金项目
国家重点基础研究发展计划(973计划)资助项目(2010CB833605) （973计划）
