Wang E D, Holland M
Shanghai Institute of Biochemistry, Chinese Academy of Sciences.
Chin J Biotechnol. 1989;5(2):73-9.
The upstream activation sequence from ENO2, one of two genes coding for yeast enolase, was inserted into the upstream 405 HpaI site of LEU2, which codes for beta-isopropylmalate dehydrogenase (E.C. 1.1.1.85), utilizing shuttle plasmid YEp13. The effect of the ENO2 upstream activation sequence on expression of yeast LEU2 was studied. Our results revealed a fourfold increase in expression for LEU2 in both orientations after activation by the ENO2 upstream activation sequence. Leucine repressed LEU2 expression. Glucose did not induce the ENO2 upstream activation sequence effect on LEU2 expression. It is possible to construct a high-level expression system in yeast by using the ENO2 upstream activation sequence.
利用穿梭质粒YEp13,将编码酵母烯醇酶的两个基因之一ENO2的上游激活序列插入到编码β-异丙基苹果酸脱氢酶(E.C. 1.1.1.85)的LEU2基因上游的405 HpaI位点。研究了ENO2上游激活序列对酵母LEU2表达的影响。我们的结果显示,在ENO2上游激活序列激活后,LEU2在两个方向上的表达均增加了四倍。亮氨酸抑制LEU2的表达。葡萄糖不会诱导ENO2上游激活序列对LEU2表达产生影响。利用ENO2上游激活序列有可能在酵母中构建一个高水平表达系统。