表达水痘-带状疱疹病毒糖蛋白E基因毕赤酵母的构建

Construction of recombinant Pichia pastoris yeast expressing glycoprotein E of the varicella-zoster virus

目的本研究拟将水痘-带状疱疹病毒(varicella-zoster virus,VZV)的主要抗原糖蛋白E在酵母表达系统中分泌表达。方法根据酵母的密码子偏爱性优化编码VZV糖蛋白E的基因后,构建真核表达载体p GAPZαA-g E,并将其转化至毕赤酵母X-33菌株中进行摇瓶表达,利用SDS-PAGE以及Western-blot鉴定其蛋白表达情况。结果经检测,在筛选得到的重组子p GAPZαA-g E/X-33摇瓶表达上清中可检测到相对分子质量约为60×103的目的蛋白条带。结论实现了水痘-带状疱疹病毒糖蛋白E基因在毕赤酵母中的分泌表达,为进一步制备水痘-带状疱疹病毒糖蛋白E、研究其免疫原性及疫苗的研发等相关性研究奠定了基础。

Objective To construct a recombinant yeast to express secretory glycoprotein E of varicella-zoster virus（VZV）in the yeast strain Pichia. Methods The DNA sequence encoding glycoprotein E of VZV was optimized for Pichia pastoris.Then,the sequence was synthesized and cloned into eukaryotic expression vector pGAPZαA.The recombinant plasmid was isolated,linearized,and transformed into the X-33 strain of the yeast Pichia pastoris by electroporation.After selection,target protein in the culture supernatant was examined by SDS-PAGE and Western blot. Results Both SDS-PAGE and Western blot results proved the expression of glycoprotein E（60×103）in the fermentation supernatant of transformants pGAPZαA-gE/X-33. Conclusions A recombinant yeast was successfully constructed to express secretory VZV-gE in the yeast strain Pichia.