金黄色葡萄球菌SaeP蛋白表达条件的优化

Optimization of Expression Conditions of Staphylococcus aureus SaeP Protein

对金黄色葡萄球菌SaeP蛋白在大肠杆菌中的表达工艺进行优化,提高SaeP蛋白的表达量,为深入研究Sae系统对金黄色葡萄球菌产生毒力因子的机制提供支持。通过单因素法以及全面实验法确定最佳的宿主菌、诱导剂浓度、诱导温度以及诱导时间。结果表明:将通过无限克隆法(Restriction Free Cloning, RF)构建的重组质粒,转入大肠杆菌BL21(DE3)表达宿主中,用浓度为0.5 mmol/L的IPTG,在16℃条件下诱导8 h,金黄色葡萄球菌SaeP蛋白的相对表达量最高金黄色葡萄球菌SaeP蛋白的最高表达量占总蛋白的35.14%,较原诱导条件提高25.49%。

Increasing the expression yield of SaeP protein from Staphylococcus aureus will benefit the research of the regulated mechanism of virulence factory and the two component regulatory system in Staphylococcus aureus. This paper optimized the expression system of SaeP protein in Escherichia coli. The results showed that the recombinant plasmid constructed by Restriction Free Cloning method, and transferred into BL21(DE3) host, then induced by 0.5 mmol/L IPTG at 16 ℃for 8 h, The expression level of SaeP protein of Staphylococcus aureus accounting for 35.14% of the cell protein, and the original condition was increased 25.49%.