军事医学2011，Vol.35Issue(1)：25-29,5.
突变型脑红蛋白原核表达载体的构建与表达鉴定
Construction and expression of a prokaryotic vector for mutant human neuroglobin
摘要
Abstract
Objective To construct the prokaryotic expression vector for expressing mutant neuroglobin(Ngb) protein in order to study the function of Ngb. Methods The mutant Ngb gene was amplified by PCR with primer-directed site-specific mutagenesis. The amplified DNA fragment was then sub-cloned into the prokaryotic expression vector pBV220. The recombinant plasmid was transferred into E. coli HB101. Expression of the mutant Ngb protein was detected with SDS-PAGE and Western blot. Results Two mutant Ngb prokaryotic expression vectors, pBV220-Ngb (H64V) and pBV220-Ngb ( H96A), were successfully constructed. The SDS-PAGE and Western blot analyses showed that the mutant Ngb proteins could be induced for expression as expected. Conclusion The preparation of the mutant Ngb protein will facilitate studies on the function as well as the mechanism for the neuroprotective effect of Ngb.关键词
脑红蛋白/神经保护/突变/原核表达/引物突变法/氨基酸类/聚合酶链反应/大肠杆菌/基因表达分类
生物科学引用本文复制引用
李伟光,吴永红,任长虹,高艳,张成岗..突变型脑红蛋白原核表达载体的构建与表达鉴定[J].军事医学,2011,35(1):25-29,5.基金项目
国家973计划项目(2006CB504100),国家科技重大专项课题(2009ZX09503-002, 2009ZX09301-002, 2009ZX09103-616), 国家自然科学基金(81070741,30973107, 30772293) （2006CB504100）
