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中国农学通报

中国农学通报 ›› 2010, Vol. 26 ›› Issue (13): 10-13.

所属专题： 生物技术； 园艺

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

耐甲氧西林金黄色葡萄球菌PBP2a蛋白转肽酶区的原核表达及鉴定

刘长浩 任慧英 刘文华 邹玲 温建新   

  • 收稿日期:2009-12-15 修回日期:2010-01-10 出版日期:2010-07-05 发布日期:2010-07-05

Prokaryotic Expression and Identification of the Transpeptidase Domain of PBP2a of Methicillin Resistant Staphylococcus aureus

  • Received:2009-12-15 Revised:2010-01-10 Online:2010-07-05 Published:2010-07-05

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摘要：

本研究对编码耐甲氧西林金黄色葡萄球菌（MRSA）非青霉素结合蛋白2a（PBP2a）的转肽酶区（TPase）原核表达并进行Western blot鉴定。首先通过PCR方法由MRSA基因组中扩增转肽酶区基因片段，并构建pGEX-6p-1-TPase重组质粒，转化BL-21，诱导目的蛋白表达，然后进行Western blot鉴定。结果表明，成功构建出pGEX-6p-1-TPase原核表达载体，并成功表达出PBP2a的转肽酶区蛋白（TPase），为其进一步的研究和应用奠定了基础。

Abstract:

The mecA fragment which encodes the transpeptidase domain of penicillin binding protein 2a(PBP2a)of methicillin resistant staphylococcus aureus(MRSA),to express and identificated the transpeptidase domain of PBP2a by western blot. The mecA fragment which encodes the transpeptidase domain of PBP2a was amplified from the genomic DNA of MRSA by PCR, and then the objective gene fragment was cloned into pGEX-6p-1 plasmid. The recombinant plasmid pGEX-6p-1-TPase was transformed into BL21.The expression was induced by IPTG and the expressed product was identificated by western blot.The results indicated that the corresponding prokaryotic expression vector was successfully constructed, and the transpeptidase domain of PBP2a was successfully expressed, which established the foundation for further investigation.