MRSA-PBP2a转肽酶区的原核表达及鉴定

Prokaryotic Expression and Identification of the Transpeptidase Domain of PBP2a of MRSA

为了对编码耐甲氧西林金黄色葡萄球菌(MRSA)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。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,经酶切鉴定、测序正确后,转化BL-21,用IPTG进行诱导表达,并对表达的蛋白进行Western blot鉴定。结果表明,构建的pGEX-6p-1-TPase原核表达载体可表达PBP2a的转肽酶区蛋白,为其进一步的研究和应用奠定了基础。

To express and identify the transpeptidase domain of penicillin binding protein 2a which was encoded by mecA fragment in methicillin resistant staphylococcus aureus（MRSA）. The mecA fragment which encodes TPase of PBP2a was amplified from the genomic DNA of MRSA by PCR, and then cloned into pGEX-6p-1 plasmid. The recombinant plasmid pGEX-6p-1-TPase was transformed into BL21 cell after identification by enzyme digestion and sequencing. The target protein was induced to express by IPTG and identificated by western-blot. The results indicated that the recombinant prokaryotic expression vector pGEX-6p-1-Tpase could produce the transpeptidase domain of PBP2a, which will lay the foundation for further investigation.