生命科学研究2012，Vol.16Issue(3)：223-227,5.
金黄色葡萄球菌甲氧西林耐药辅助基因femB的克隆和原核表达
Cloning and Prokaryotic Expression of femB Gene of Staphylococcus aureus
摘要
Abstract
FemB gene is closely related with high level of methicillin resistance of Staphylococcus aureus(S. aureus) and is a potential new target for antibiotics against MRSA. Genomic DNA from S. aureus was used as template for PCR amplification of femB gene. The obtained femB gene fragment was cloned into pGM-T vector and then transformed into the competent E. coli DH5a. The positively clone was identified by PCR, restriction endonuclease analysis and DNA sequencing. The identified fragment was cloned into expression vector pGEX-4T-l and transformed into E. coli BL21, and then induced by IPTG to express GST/FemB fusion protein. The expressed protein was identified by SDS-PAGE and Western blot. Results showed that femB gene was cloned and recombinant plasmid was constructed successfully. The recombinant protein with a relative molecular mass 75 kD, which specifically combined with the anti-GST-tag antibody, was highly expressed in E.coli. A prokaryotic expression system of femB gene has been constructed successfully, which laid foundations for investigation on its biological functions.关键词
金黄色葡萄球菌/femB基因/基因克隆/融合蛋白Key words
Staphylococcus aureus/ femB gene/ gene clone/ fusion protein分类
医药卫生引用本文复制引用
邹明祥,武文君,李军,邬国军,张宁洁,刘文恩,范学工..金黄色葡萄球菌甲氧西林耐药辅助基因femB的克隆和原核表达[J].生命科学研究,2012,16(3):223-227,5.基金项目
湖南省自然科学基金资助项目(10JJ5027) （10JJ5027）
中南大学本科生自由探索研究创新基金资助项目(2010112001166) （2010112001166）
