重组溶葡萄球菌蛋白的原核表达、纯化及生物学活性研究

Prokaryotic expression,purification of recombinant lysostaphic protein and an initial study on its biological activities

目的原核表达重组溶葡萄球菌蛋白并检测其杀菌效力。方法在成熟溶葡萄球菌蛋白编码序列的基础上,设计重组序列,采用PCR技术扩增,克隆至大肠埃希菌表达载体,IPTG诱导表达,用微量稀释法研究其对金黄色葡萄球菌(简称金葡菌)的杀菌效果。结果目的DNA片段克隆至表达载体后经测序鉴定正确,在大肠埃希菌中获得高效表达,经免疫印迹鉴定重组蛋白表达正确。重组溶葡萄球菌蛋白对金葡菌的最小抑菌浓度(minimum inhibitory concentration,MIC)和最小杀菌浓度(minimum bactericidal concentration,MBC)均小于传统抗生素。结论成功表达重组溶葡萄球菌蛋白,其对金葡菌有强大的杀菌效力。

Objective To express lysostaphic protein in Escherichia coli logical activities. Methods The coding sequence of mature protein obtained （E. coli. ） and evaluate its bioby PCR was cloned into the prokaryotic expression plasmid. The target protein was expressed in E. coli. induced by IPTG. The bacteriocidal activities of purified protein were evaluated in vitro by microdilution method. Results The recombinant plasmid was identified by DNA sequencing. The protein was successfully expressed in E. coli. and verified by Western blotting. The MIC and MBC of the protein to staphylococcus aureus were smaller than the traditional antibiotics. Conclusion We have successfully expressed competent lysostaphic protein in prokaryotic expression system, which has a powerful bactericidal ability against staphylococcus aureus.