Wen D Z, Schlesinger M J
Proc Natl Acad Sci U S A. 1986 Jun;83(11):3639-43. doi: 10.1073/pnas.83.11.3639.
cDNAs encoding either the structural proteins (capsid and glycoproteins E1 and E2) of Sindbis virus or the glycoprotein of vesicular stomatitis virus (VSV) were fused to the Saccharomyces cerevisiae galactokinase gene (GAL1) promoter and inserted into a yeast shuttle vector. After addition of galactose to yeast transformed with this vector, 2.5-3% of total yeast protein synthesis was detected as virus proteins by specific anti-virus protein antibodies. In cells containing the Sindbis virus structural genes, the virus capsid protein was effectively released from the nascent polypeptide and two endoglycosidase H-sensitive glycoproteins were produced. One of these was identical in its gel mobility to E1 and the other appeared to be p62, a precursor to E2. A low level of E1 protein was detected on the cell's surface membranes. A single molecular weight species of glycosylated VSV glycoprotein was produced and half of the total protein could be detected at the surface membranes of yeast. Addition of long mannose chains and acylation of the virus proteins with fatty acids were not observed. Formation of virus proteins was also examined in yeast secretory mutants; one of these (sec53) failed to glycosylate the virus proteins.
编码辛德毕斯病毒结构蛋白（衣壳蛋白以及糖蛋白E1和E2）或水泡性口炎病毒（VSV）糖蛋白的cDNA与酿酒酵母半乳糖激酶基因（GAL1）启动子融合，并插入酵母穿梭载体。在用该载体转化的酵母中加入半乳糖后，用特异性抗病毒蛋白抗体检测到，总酵母蛋白合成的2.5 - 3%为病毒蛋白。在含有辛德毕斯病毒结构基因的细胞中，病毒衣壳蛋白从新生多肽中有效释放出来，并产生了两种对内切糖苷酶H敏感的糖蛋白。其中一种在凝胶迁移率上与E1相同，另一种似乎是p62，即E2的前体。在细胞表面膜上检测到低水平的E1蛋白。产生了单一分子量的糖基化VSV糖蛋白，并且在酵母表面膜上可检测到总蛋白的一半。未观察到病毒蛋白添加长甘露糖链以及脂肪酸酰化的情况。还在酵母分泌突变体中检测了病毒蛋白的形成；其中一个突变体（sec53）无法对病毒蛋白进行糖基化。
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