金黄色葡萄球菌肠毒素C的原核表达、纯化及活性鉴定

Expression,Purification and Superantigen Activity Identification of Staphylococcus aureus Enterotoxin C

目的:在大肠杆菌中表达金黄色葡萄球菌肠毒素C(SEC),并对其活性进行检测。方法:以金黄色葡萄球菌基因组为模板扩增SEC基因,经测序正确后插入原核表达载体pBV220中,转化大肠杆菌DH5α后温度热诱导重组SEC表达;表达产物经阳离子柱纯化后,用ELISA鉴定其抗原性;通过观测表达产物对鼠脾淋巴细胞的刺激增殖情况,检测其超抗原活性。结果:构建了SEC-pBV220原核表达载体,在大肠杆菌中可快速、高效表达SEC蛋白,表达产物具有超抗原活性。结论:实现了SEC的原核表达。

Objective：To express the Staphylococcus aureus enterotoxin C（SEC） in E.coli and to analyse its superantigen activity.Methods：The gene sequence of SEC was amplified by PCR using Staphylococcus aureus genome as template.The PCR product was confirmed by sequence analysis.And then SEC-pBV220 was constructed and transformed into E.coli DH5α,and recombinant SEC protein was expressed by temperature induction and purified by CM ion-exchange chromatography.ELISA was performed to detect its immunogenicity,and the proliferation of mouse spleen lymphocyte was used to test its superantigen activity.Results：The prokaryotic expression vector SEC-pBV220 was constructed and SEC was expressed in E.coli efficiently.The expressed protein has the superantigen activity after purification.Conclusion：The protein SEC could be successfully expressed in prokaryotic expression system.