Jigami Y, Toshimitsu N, Fujisawa H, Uemura H, Tanaka H, Nakasato S
J Biochem. 1986 Apr;99(4):1111-25. doi: 10.1093/oxfordjournals.jbchem.a135575.
Yeast ENO1 promoter was prepared by a chemical synthetic method. Two variant promoters containing a pyrimidine-rich region (CT block), located between the TATA box and the transcription start site, either 32 or 34 base pairs (bp) longer than the native ENO1 promoter were isolated during the chemical synthesis. Gene expression of variant promoters was compared with that of the native promoter by measuring the amount of mRNA and the activity of beta-galactosidase by constructing ENO1-lacZ gene fusions. No significant differences were observed between the native and variant promoters in transcription levels. The start site of transcription was mapped on CAAG, a consensus sequence of transcription start site of yeast glycolytic genes. The results suggest a longer CT block in ENO1 promoter may not affect the expression of the yeast ENO1 gene. In addition, the level of ENO1 gene expression was found to be higher in stationary phase cells than in log phase cells.
酵母ENO1启动子通过化学合成方法制备。在化学合成过程中分离出了两个变体启动子，它们在TATA框和转录起始位点之间含有一个富含嘧啶的区域（CT块），比天然ENO1启动子长32或34个碱基对（bp）。通过构建ENO1-lacZ基因融合体，通过测量mRNA的量和β-半乳糖苷酶的活性，将变体启动子的基因表达与天然启动子进行了比较。在转录水平上，天然启动子和变体启动子之间未观察到显著差异。转录起始位点定位于CAAG，这是酵母糖酵解基因转录起始位点的共有序列。结果表明，ENO1启动子中较长的CT块可能不会影响酵母ENO1基因的表达。此外，发现静止期细胞中ENO1基因的表达水平高于对数期细胞。
