一种具有磷酸酶活性的金黄色葡萄球菌蛋白的初步鉴定

Elementary Identification of Activity of Phosphatase of Protein from Staphylococcus aureus

金黄色葡萄球菌是一种人畜重要的致病微生物,寻找新型药物靶标是及时有效治疗金黄色葡萄球菌感染的潜在途径。本实验依据数据库,体外设计并合成了一对特异引物,应用PCR方法扩增出一种新的磷酸酶基因,并将该基因克隆至表达载体p ET2a上。经测序、酶切和PCR鉴定正确后转化大肠杆菌感受态细胞BL21中,经IPTG体外诱导表达出重组蛋白并对其进行了磷酸酶活性测定。结果表明:该基因编码蛋白具有更多依赖于镁离子的磷酸酶活性,这种酶活性受到特异性抑制剂氟化钠的抑制。研究结果为进一步挖掘该蛋白作为金黄色葡萄球菌感染有效治疗药物靶点提供了数据资料。

Staphylococcus aureus is an important pathogenic microorganism,and the exploration of novel drug tar-gets is a potential way to effective timely conduct the infection of Staphylococcus aureus.Based on the database,a pair of special primers were designed and synthesized in vitro.A new phosphatase gene was amplified by PCR,and the target gene was cloned to the vector pET2a.After sequencing,enzyme digestion and PCR identification,the re-combinant plasmid was transformed into Escherichia coli BL21.The recombinant protein was induced by IPTG in vi-tro and the activity of enzyme was determined.The results showed that the protein coded by the target gene had activ-ity of phosphatase which was more dependent on magnesium ion.The activity of protein enzyme was specific inhibit-ed by sodium fluoride.The results provide data for the further excavation of this protein as an effective therapeutic drug target for Staphylococcus aureus infection.