Silva A, Benítez J
Facultad de Biología, Universidad de La Habana, Cuba.
Arch Med Res. 1994 Winter;25(4):447-9.
To develop a yeast system that expresses bacterial and viral antigens in the future, we analyzed yeast secretion with a reporter system based on the cc1A gene of Clostridium thermocellum that encodes endoglucanase A. Sequences coding for the mature CelA protein were fused in frame to yeast promoters and secretion signals, introduced into different vectors and analyzed biochemically in yeast secretory mutants. Gene dosage had little effect, whereas expression from a weak promoter enhanced endoglucanase A secretion. In yeast secretory mutants, a major fraction of endoglucanase A piled up in the endoplasmic reticulum as a core glycosylated protein.
为了开发一个未来能够表达细菌和病毒抗原的酵母系统,我们使用了一个基于热纤梭菌cc1A基因(编码内切葡聚糖酶A)的报告系统来分析酵母分泌情况。编码成熟CelA蛋白的序列与酵母启动子和分泌信号框内融合,导入不同载体,并在酵母分泌突变体中进行生化分析。基因剂量影响不大,而来自弱启动子的表达增强了内切葡聚糖酶A的分泌。在酵母分泌突变体中,大部分内切葡聚糖酶A作为核心糖基化蛋白在内质网中堆积。