登革病毒衣壳蛋白与葡萄球菌核酸酶融合蛋白在大肠杆菌中的表达  

Expression of the fusion protein of dengue virus capsid protein C and staphylococcal nuclease in Escherichia coli

作　　者：秦成峰[1] 胡志君[1] 陈水平[1] 范宝昌[1] 于曼[1] 姜涛[1] 邓永强[1] 段鸿元[1] 秦鄂德[1] 

机构地区：[1]军事医学科学院微生物流行病研究所,北京100071

出　　处：《军事医学科学院院刊》2004年第5期430-432,491,共4页Bulletin of the Academy of Military Medical Sciences

基　　金：国家自然科学基金资助课题 (3 0 10 0 0 0 6)

摘　　要：目的 :实现登革病毒衣壳蛋白C与葡萄球菌核酸酶SN融合蛋白在大肠杆菌中的表达。方法 :利用基因重组技术将通过BamHⅠ连接在一起的CSN基因克隆入表达载体 pLEX ,转化大肠杆菌GI72 4并以色氨酸诱导表达 ,用SDS PAGE和免疫印迹法鉴定表达的融合蛋白 ,采用TDA显色法检测融合蛋白中SN的活性。结果与结论 :成功构建重组表达载体pLEX CSN并在大肠杆菌中获得表达 ,表达的融合蛋白相对分子质量约 2 70 0 0 ,可被抗登革病毒C蛋白抗体识别 ,并具有SN的生物活性。为探讨登革病毒衣壳蛋白靶向性抗病毒作用奠定了基础。Objective: To express the fusion protein of dengue vi rus capsid(DEN) C and staphylococcal nuclease in Escherichia coli. Methods: The DEN C gene was ligated with SN gene at BamHⅠ site. The fusion gene was then subcloned into the pLEX vector and expressed in E.coli GI724. The recombinant fusion protein was identified by SDS-PAGE and Western blot, and the SN was characterized by TDA method. Results and C onclusions: The fusion protein was effectively expressed. Its relativ e molecular weight was about 27 000. The DEN C protein in the fusion protein was confirmed by Western blot, and the SN was enzymatically active.

关 键 词：登革病毒 衣壳蛋白 葡萄球菌核酸酶 融合蛋白 表达 

分 类 号：R346[医药卫生—基础医学]