Feng B, Li Y Y, Chen Z C
Department of Molecular Biology, Tongji Medical University, Wuhan.
J Tongji Med Univ. 1993;13(2):77-83. doi: 10.1007/BF02887920.
The plasmid series YEP51 delta n bearing GAL10-SUC2 promoter combinations were constructed by inserting SUC2 gene with different upstream deletions downstream GAL10 promoter on YEP51. After transforming yeast cells S. cerevisiae, the invertases expressed by each of the transformants were measured and analysed by means of PAGE. The results showed that: 1) The SUC2 gene with upstream deletion to at -636bp expressed high level glycosylated form of invertase under glucose derepression, while SUC2 gene with more extensive deletions to -223 bp or more lost its response to glucose derepression; 2) Each part of GAL10-SUC2 promoter combination acted almost independently. All of the combinations showed no apparent coordinated promoter function under our experimental conditions; 3) Sequences between -89bp and -41bp of SUC2 upstream region are responsible for constitutive expression of nonglycosylated invertase. The two tracts of poly (dA-dT) of this region may serve as promoter elements.
通过将带有不同上游缺失的SUC2基因插入YEP51上的GAL10启动子下游，构建了携带GAL10 - SUC2启动子组合的YEP51δn质粒系列。将酵母细胞酿酒酵母转化后，通过PAGE对每个转化体表达的转化酶进行了测定和分析。结果表明：1）上游缺失至 - 636bp的SUC2基因在葡萄糖去阻遏条件下表达高水平糖基化形式的转化酶，而缺失至 - 223bp或更多的更广泛缺失的SUC2基因失去了对葡萄糖去阻遏的反应；2）GAL10 - SUC2启动子组合的每个部分几乎独立起作用。在我们的实验条件下，所有组合均未显示出明显的协同启动子功能；3）SUC2上游区域 - 89bp至 - 41bp之间的序列负责非糖基化转化酶的组成型表达。该区域的两条聚（dA - dT）序列可能作为启动子元件。
