【摘要】 目的构建含SEA基因的原核表达载体,并在大肠杆菌表达系统中表达。方法应用PCR扩增SEA基因片段,与克隆载体pGEMT-easy连接,插入到表达载体pET-30a中,转化大肠杆菌BL21(DE3),经诱导表达及鉴定。结果PCR扩增约770bp的基因片段,克隆至载体后,经测序与文献报道结果一致,表达蛋白相对分子质量约31000,纯化后鉴定为SEA蛋白。结论已成功获得SEA蛋白,为其进一步研究和应用奠定基础。更多还原

【Abstract】 Objective To construct the prokaryotic expression vector of staphylococcal enterotoxin A(SEA) gene and express the gene in E.coli.Methods Amplify SEA gene by PCR,ligate to cloning vector pGEM T-easy,then insert into expression vector pET-30a and transform to E.coli for expression.Results A gene fragment at a length of 770 bp was amplified,and its sequence was identical to that reported.The protein with a relative molecular weight of 31 000 was expressed and identified as SEA after purification.Conclusion SEA protein was successfully expressed.It laid a foundation of further study and application of superantigen SEA.更多还原