D2-43病毒E蛋白在酵母细胞表面的展示

目的：在酵母细胞表面展示登革2型病毒43株(D2—43)的E基因，探索利用酵母表面展示系统建立DNA改组筛选平台的可行性。方法：通过RT-PCR扩增获得D2-43的E基因，将该基因亚克隆至T载体后，再克隆至酵母表面展示载体pYDI，于酿酒酵母EBY100中利用半乳糖进行诱导表达。表达产物采用间接免疫荧光法和FACS进行检测。结果：酵母表面展示产物可与D2-43的腹水抗体特异性地结合；在半乳糖诱导后24h，展示E蛋白的酵母细胞百分数达22．07％。结论：本研究为建立基于酵母表面展示系统的DNA改组筛选平台奠定了基础。

Display of the envelope protein of D2-43 on yeast cells surface

Objective:To display the envelope protein of dengue virus type 2 strain 43 (D2-43) on yeast cell surface and explore a screening platform for DNA shuffling with yeast surface display system. Methods:The envelope gene of D2-43, which was amplified with RT-PCR, was subcloned into T vector, and then cloned into the yeast surface display vector pYD1.Epression was induced with galactose in Saccharomyces cerevisiae EBY100. Displayed products were detected with indirect immunofluorescence assay and FACS.Results:Displayed products on yeast cell surface interacted specifically with ascitic antibody against D2-43 and the percentage of yeast cells displaying the envelope protein was 22.07% after yeast cells were induced with galactose for 24 hours. Conclusion:The study lays a foundation to establish a screening platform for DNA shuffling based on yeast surface display system.