现代检验医学杂志2012，Vol.27Issue(5)：57-60,4.DOI:10.3969/j.issn.1671-7414.2012.05.019
耐甲氧西林金黄色葡萄球菌PBP2a的真核表达及其在抗体鉴定中的应用
Eukaryotic Expression of PBP2a in Methicillin Resistant Staphylococcus Aureus and Application in Antibody Identification
摘要
Abstract
Objective To express methicilin resistant Staphylococcus aureus penicillin binding protein 2a (PBP2a) with bacu-lovirus expression system. Methods MecA gene was obtained by double digest pGEMT-mecA plasmid using EcoR I and BamH I, and then the gene was inserted into plasmid pFastBacl. Recombinant plasmid was transformed into E. Coli DHlOBac containing a shuttle vector,Bacmid. Recombinant shuttle vector bacmid-mecA was obtained by site-specific transposition and transfected into sf9 cells. The expressed protein was identified by IFA and Western blot analysis. Results The baculovirus which containing mecA gene was expressed successfully. Morphological changes displayed in the transfected sf9 cells assumed that the transfection was successful. IFA and Western blot proved the expressed protein could combine with PBP2a polyclonal antibody. Conclusion The PBP2a protein was expressed successfully by Bac-to-Bac baculovirus expression system.关键词
耐甲氧西林金黄色葡萄球菌/青霉素结合蛋白2a/真核表达Key words
methicillin resistant staphylococcus aitreus/PBP2a/ eukaryotic expression分类
医药卫生引用本文复制引用
邹玉涵,朱祖怀,闫佩毅,朱元,黄德魁,朱晴晖,金姝..耐甲氧西林金黄色葡萄球菌PBP2a的真核表达及其在抗体鉴定中的应用[J].现代检验医学杂志,2012,27(5):57-60,4.基金项目
上海市普陀区科技自主创新资助项目(PTKW08-B01),普陀区卫生系统自主创新科研资助项目(普KW11103). （PTKW08-B01）
