摘要
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.
出处
《生物技术通讯》
CAS
2008年第4期512-514,共3页
Letters in Biotechnology
基金
全军"十一五"重点专题(06Z069)
关键词
金黄色葡萄球菌肠毒素C
原核表达
活性
Staphylococcus aureus enterotoxin C
prokaryotic expression
superantigen activity