Li W, Li Y
Institute of Genetics, Fudan University, Shanghai.
Yi Chuan Xue Bao. 1997;24(6):561-8.
By fusing the upstream activation sequence (UAS) of ADH2 gene to the 5' end of a series of deletions in the upstream region of SUC2 gene, 4 expression plasmids YFD110 delta 1, YFD110 delta 9, YFD110 delta 17 and YFD110 delta 11 were constructed. They contained different hybrid promoters for transcription of SUC2 gene. After transforming them together with two control plasmids YFD26 delta 1, YFD25 into yeast. S. cerevisiae Y33 respectively, the transformants were grown in the repression or derepression media and the invertase produced by each transformants were analyzed by colormetry and gel electrophoresis. The results were as follow is: (1) UASSUC2 and UASADH2 in the hybrid plasmid YFD110 delta 1 worked synergically under derepression condition. Under repression condition, Y33/YFD110 delta 1 produced very low level of glycosylation invertase. (2) Compared with three different derepression media, the medium containing low concentration glucose gave higher derepression efficiency of glycosylated invertase than the media containing glycerol and ethanol.
通过将ADH2基因的上游激活序列（UAS）融合到SUC2基因上游区域一系列缺失片段的5'端，构建了4种表达质粒YFD110δ1、YFD110δ9、YFD110δ17和YFD110δ11。它们含有用于SUC2基因转录的不同杂合启动子。将它们与两种对照质粒YFD26δ1、YFD25一起分别转化酿酒酵母Y33后，将转化体在阻遏或去阻遏培养基中培养，并通过比色法和凝胶电泳分析每个转化体产生的转化酶。结果如下：（1）杂交质粒YFD110δ1中的UASSUC2和UASADH2在去阻遏条件下协同作用。在阻遏条件下，Y33/YFD110δ1产生的糖基化转化酶水平非常低。（2）与三种不同的去阻遏培养基相比，含低浓度葡萄糖的培养基比含甘油和乙醇的培养基对糖基化转化酶的去阻遏效率更高。
