Uemura H, Shiba T, Paterson M, Jigami Y, Tanaka H
Gene. 1986;45(1):67-75. doi: 10.1016/0378-1119(86)90133-2.
A DNA fragment which contains the 5'-flanking region of the Saccharomyces cerevisiae enolase 1 gene (ENO1) and a portion of the coding sequence was cloned in a plasmid pMC1587. This fragment was fused in frame to the lacZ gene of Escherichia coli. Many mutants which deleted a portion of the 5'-flanking region of ENO1 were isolated from this ENO1-lacZ fusion plasmid by in vitro recombination. Analysis of beta-galactosidase activity of these mutants indicated that the regulatory region responsible for an efficient expression of the ENO1-lacZ fused gene resides within an 86-bp sequence located at -487 to -402 upstream from the start codon of ENO1. We found that the segment encompassing the 86-bp region worked equally well in an inverted orientation, but the tandem duplication of the sequence did not enhance the expression of the fused gene.
一个包含酿酒酵母烯醇化酶1基因（ENO1）5'侧翼区和部分编码序列的DNA片段被克隆到质粒pMC1587中。该片段与大肠杆菌的lacZ基因读框融合。通过体外重组从这个ENO1 - lacZ融合质粒中分离出许多缺失ENO1 5'侧翼区一部分的突变体。对这些突变体的β-半乳糖苷酶活性分析表明，负责ENO1 - lacZ融合基因高效表达的调控区位于ENO1起始密码子上游-487至-402处的一个86 bp序列内。我们发现，包含86 bp区域的片段以反向排列时同样有效，但该序列的串联重复并没有增强融合基因的表达。
