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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 achieve the expression of Dengue virus capsid protein C and staphylococcal nuclease SN fusion protein in Escherichia coli. Methods: The CSN gene linked by BamHⅠ gene was cloned into the expression vector pLEX by gene recombination technology, transformed into E. coli GI72 4 and induced by tryptophan. The expressed fusion protein was identified by SDS PAGE and Western blot. Colorimetric assay of SN activity in fusion proteins. RESULTS AND CONCLUSION: The recombinant expression vector pLEX CSN was successfully constructed and expressed in E. coli. The expressed fusion protein has a relative molecular mass of about 2 700 and can be recognized by anti-dengue virus C protein antibody and has the biological activity of SN. Which laid the foundation for the study of the targeted antiviral effect of dengue virus capsid protein.