Istituto Biosintesi Vegetali, C.N.R., Via Bassini, 15, 20133 Milano, Italy.
EMBO J. 1986 Mar;5(3):459-65. doi: 10.1002/j.1460-2075.1986.tb04234.x.
Maize genomic fragments containing the regulatory and coding regions of a zein gene for a low size class 23-kd protein have been inserted in an interspecific Escherichia coli-Saccharomyces cerevisiae expression vector in different constructions. The presence of the inducible GAL1-10 upstream activation site (UAS) allows us to regulate differentially by carbon sources the transcription of the zein gene both under the plant promoter and under the yeast CYC-1 promoter. We found that the zein promoter region is properly recognized at the correct transcription start, while different termination points occur during transcription. The yeast UAS was also shown to function as a typical eukaryotic enhancer regardless of its distance or orientation with respect to the plant promoter. Yeast cells transformed by a plasmid containing a zein sequence fused to a short piece of the CYC-1 gene produced a fused polypeptide, of expected mol. wt, in variable amount from 0.2 to 5% depending on the growth phase conditions.
已经将包含低分子量 23kD 蛋白的 zein 基因调控区和编码区的玉米基因组片段插入到不同构建体的种间大肠杆菌-酿酒酵母表达载体中。诱导型 GAL1-10 上游激活序列 (UAS) 的存在使我们能够通过碳源对 zein 基因的转录进行差异调控,无论是在植物启动子还是在酵母 CYC-1 启动子的控制下。我们发现 zein 启动子区域在正确的转录起始处被正确识别,而在转录过程中会发生不同的终止点。酵母 UAS 也被证明是一种典型的真核增强子,无论其与植物启动子的距离或方向如何。含有 zein 序列与短段 CYC-1 基因融合的质粒转化的酵母细胞产生了一种预期分子量的融合多肽,其产量在 0.2%到 5%之间变化,具体取决于生长阶段的条件。