Objectives

To explore Saccharomyces cerevisiae as an expression platform for dengue oral immune complex vaccine development.

Results

Molecular engineering was applied to create a fusion gene construct (scEDIII-PIGS) consisting of a yeast codon optimized sequence encoding for a synthetic consensus dengue envelope domain III (scEDIII) followed by a modified IgG Fc domain (PIGS). Northern blot showed transcription of the target gene, with a temporal expression pattern similar to those from previous work. Western blot showed assembly of various immune complexes from monomer to hexamer. Partial purification of scEDIII-PIGS was also attempted to demonstrate the feasibility of yeast system for immune complex vaccine development. Approximately 1 mg of scEDIII-PIGS can be produced from 1 l culture.

Conclusion

This work demonstrated for the first time that various immunocomplex structures of our target protein could be efficiently produced in S. cerevisiae for future application in developing oral and injectable vaccines against various pathogens.

中文翻译：

---

由与共有登革热包膜结构域 III 融合的 Fc 片段组成的免疫复合物在酿酒酵母中的表达

目标

探索酿酒酵母作为登革热口服免疫复合物疫苗开发的表达平台。

结果

应用分子工程来创建融合基因构建体 (scEDIII-PIGS)，该构建体由酵母密码子优化的序列组成，该序列编码合成的共有登革热包膜域 III (scEDIII)，然后是修饰的 IgG Fc 域 (PIGS)。Northern印迹显示靶基因的转录，其时间表达模式与以前的工作相似。蛋白质印迹显示从单体到六聚体的各种免疫复合物的组装。还尝试对 scEDIII-PIGS 进行部分纯化，以证明酵母系统用于免疫复合物疫苗开发的可行性。1 l 培养物可以产生大约 1 mg scEDIII-PIGS。

结论

这项工作首次证明，我们的靶蛋白的各种免疫复合物结构可以在酿酒酵母中有效产生，用于未来开发针对各种病原体的口服和注射疫苗。