Mokdad-Gargouri Raja, Abdelmoula-Soussi Salma, Hadiji-Abbès Nadia, Amor Ines Yacoubi-Hadj, Borchani-Chabchoub Istabrak, Gargouri Ali
Centre de Biotechnologie de Sfax, Sfax, Tunisia.
Methods Mol Biol. 2012;824:359-70. doi: 10.1007/978-1-61779-433-9_18.
The yeasts Saccharomyces cerevisiae and Pichia pastoris are attractive hosts for production of human proteins. The main advantages offered by these systems are the well-developed and easily accessible genetic tools, rapid growth, the simple and inexpensive culture media, and many of the cellular and metabolic processes found in higher eukaryotes are conserved in both yeast species. In this chapter, we describe the production of two proteins of therapeutic interest: the human P53 tumor suppressor and the viral HBsAg in P. pastoris and S. cerevisiae using the strong and inducible promoters AOX1 and Gal10/Cyc1, respectively. Besides the production as a goal of both expressions, we also report on an unexpected result that has occurred in S. cerevisiae: The overexpression of human p53 induces yeast cell death with characteristic markers of apoptosis, such as the externalization of phosphatidylserines and DNA strand cleavage.
酿酒酵母和巴斯德毕赤酵母是生产人类蛋白质的理想宿主。这些系统的主要优点包括：遗传工具完善且易于获取、生长迅速、培养基简单廉价，并且高等真核生物中许多细胞和代谢过程在这两种酵母中都保守存在。在本章中，我们描述了利用分别为强诱导型启动子AOX1和Gal10/Cyc1，在巴斯德毕赤酵母和酿酒酵母中生产两种具有治疗意义的蛋白质：人类p53肿瘤抑制蛋白和病毒乙肝表面抗原（HBsAg）。除了将蛋白质生产作为两种表达的目标外，我们还报告了在酿酒酵母中出现的一个意外结果：人类p53的过表达会诱导酵母细胞死亡，并伴有凋亡的特征性标记，如磷脂酰丝氨酸外化和DNA链断裂。
