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Prokaryotic expression of PBP2a of methicillin resistant staphylococcus aureus and preparation of its polyclonal antibody

Jin Shu1,Zhu Zuhuai1,Yan Peiyi1,Zou Yuhan1,Zhu Yuan1,Huang Dekui2,Zhu Qinghui1 (1.Department of Clinical Laboratory;2.Department of Cardiovascular Medicine,Shanghai People′s Hospital of Putuo District,200060,China)  
Objective To express the PBP2a of methicillin resistant staphylococcus aureus,and prepare its polyclonal antibody.Methods According to the sequence of mecA gene published on GenBank,primers were designed and synthesized.The full length of mecA gene was obtained by PCR,and then the gene was cloned into pET32a(+)plasmid,and the recombinant plasmid was transformed into E.coli BL21(DE3).The expression was induced by IPTG and the expressed product was purified by Ni2+ affinity chromatography.Mice were immuned with the purified protein for 3 to 8 times,and mice serum was collected for identification.Results The recombinant expression vector for PBP2a was constructed successfully,and PBP2a protein was highly expressed in prokaryotic expression system.The purified protein was used to prepare polyclonal antibody.Conclusion Using molecular cloning technology,purified PBP2a protein can be obtained and prepare its polyclonal antibody,which establishes the foundation for further investigation.
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