Padkina M V, Parfenova L V, Gradoboeva A E, Sambuk E V
Prikl Biokhim Mikrobiol. 2010 Jul-Aug;46(4):448-55.
The HuIFNA16, HuIFNB, and BoIFNG genes encoding human [alpha]16, beta-interferons and bovine gamma-interferon were cloned under the control of the yeast Pichia pastoris AOX1 gene promoter. The yeast strains producing heterologous interferons intracellularly and extracellularly were constructed. There was no effect of high level of heterologous protein synthesis on the yeast P. pastoris cell growth, unlike yeast Saccharomyces cerevisiae. The considerable part of the heterologous interferons was detected in the yeast P. pastoris soluble protein fraction but not in the "inclusion bodies." The treatment of human beta-interferon with endoglycosidase H showed that protein was expressed in glycosilated and unglycosilated forms. On the strength of these data, the hypothesis was suggested that the more effective heterologous gene expression in yeast P. pastoris and enhanced resistance of the methylotrophic yeast to negative effects of recombinant proteins was due to the special features of its metabolism.
编码人α16、β干扰素和牛γ干扰素的HuIFNA16、HuIFNB和BoIFNG基因在酵母巴斯德毕赤酵母AOX1基因启动子的控制下进行克隆。构建了在细胞内和细胞外产生异源干扰素的酵母菌株。与酿酒酵母不同，高水平的异源蛋白合成对巴斯德毕赤酵母细胞生长没有影响。在巴斯德毕赤酵母可溶性蛋白组分中检测到相当一部分异源干扰素，但在“包涵体”中未检测到。用内切糖苷酶H处理人β干扰素表明，该蛋白以糖基化和非糖基化形式表达。基于这些数据，提出了一个假设，即异源基因在巴斯德毕赤酵母中更有效的表达以及甲基营养型酵母对重组蛋白负面影响的增强抗性是由于其代谢的特殊特征。
