神经损伤与功能重建2012，Vol.7Issue(4)：238-240,262,4.DOI:10.3870/sjsscj.2012.04.002
哺乳动物极性蛋白mInscuteable-C末端肽段/GST融合蛋白的原核表达与纯化
Construction and Purification of a Prokaryotic Expression Vector of GST-mInsc257 ～ 532 Fusion Protein
摘要
Abstract
Objective: To construct a prokaryotic expression vector of GST-mInsc257 ～ 532 fusion protein and to express and purify the fusion protein in E. coli. Methods: A prior construc ted mlnscuteable fragment, the coding sequence of the C terminal of mlnscuteable including the 257 ～ 532 amino acids, was cloned into pGEX-4T vector down stream of the GST tag to construct a recombinant plasmid pGEX-4T/mInsc257 ～ 532. Then the plasmid was transformed into E. col/BL21 and induced to express fusion protein GST/mInsc257～532 with IPTG. The recombi nant GST-mInsc257～532 fusion protein was purified by Glutathione Sepharose beads and then analyzed by SDS-PAGE and western blotting. Results: The highly expressed and purified pGEX- 4T/mInsc257～532 recombinant fusion protein was successfully obtained. Conclusion; The re combinant plasmid pGEX-4T/mInsc257 ～ 532 was successfully constructed. GST/mInsc257 ～ 532 fusion protein was successfully expressed and purified.关键词
mInscuteable/GST/融合蛋白/原核表达/蛋白纯化Key words
mlnscuteable/ GST/ fusion protein/ prokaryotic expression/ protein purifica-tion分类
医药卫生引用本文复制引用
肖卓妮,徐望明,杨菁..哺乳动物极性蛋白mInscuteable-C末端肽段/GST融合蛋白的原核表达与纯化[J].神经损伤与功能重建,2012,7(4):238-240,262,4.基金项目
国家自然科学基金(No.81100418) （No.81100418）
