金黄色葡萄球菌GapC基因的克隆和表达  

Cloning and Expression of the Gene of Staphylococcus aureus GapC

作　　者：张晶[1] 朱洪伟[1,2] 崔玉东[1] 

机构地区：[1]黑龙江八一农垦大学,大庆163319 [2]中国农业科学院哈尔滨兽医研究所

出　　处：《黑龙江八一农垦大学学报》2008年第3期50-53,共4页journal of heilongjiang bayi agricultural university

摘　　要：为研究金黄色葡萄球菌(Staphylococcus aureus,S.aureus)GapC蛋白,应用PCR方法扩增出S.aureus菌株BMSA/855/23-1的gapC基因,随后将其克隆到pMD18-T载体上。重组克隆pMD-T/gapC经测序后,与GenBank上已发表的序列进行对比分析。然后,将gapC基因插入到pQE-30质粒,构建重组质粒pQE30/gapC。重组质粒转化大肠杆菌E.coli M15(pREP4),IPTG诱导后,SDS-PAGE鉴定GapC融合蛋白的表达。结果表明成功扩增并克隆了S.aureus gapC,该菌株的gapC基因与GenBank上S.aureus BM10株的核苷酸序列一致性为99.3%,推导氨基酸序列一致性为99.4%。SDS-PAGE结果表明,GapC蛋白在E.coli M15(pREP4)中获得表达。In order to characterize the Staphylococcus aureus （S. aureus） surface protein GapC, gapC gene of S. aureus strain BMSM 855/23-1 was amplified by PCR and was introduced into pMD18-T vector subsequently. The recombinant clone, pMD-T/gapC, was se- quenced. The nucleotide sequence of gapC was compared and analyzed with the one published previously in GenBank, and then, pQE30/gapC recombinant plasmid was constructed by inserting gapC gene into pQE-30 vector. The recombinant plasmid was trans- formed into E. coli strain M15 （pREP4）, and was induced with IPTG to express recombinant GapC fusion proein. The expression of target protein was detected by SDS-PAGE. The results showed that gapC gene of S. aureus strain BMSM855/23-1 was successfully amplified and cloned. Sequencing results showed that the gapC gene of the isolated strain shares 99.3% homology in nucleotide sequence, and 99.4% in amino acid sequence with that of the S. aureus strain BM10 in GenBank. SDS-PAGE result showed that the recombinant GapC was expressed successfully in E. coli M 15（pREP4） strain.

关 键 词：金黄色葡萄球菌 GapC蛋白 克隆 表达 

分 类 号：Q786[生物学—分子生物学]