Iborra F, Raynal A, Guerineau M
Laboratoire de Biologie et Génétique Moléculaire, Université Paris-Sud, Orsay, France.
Mol Gen Genet. 1988 Jul;213(1):150-4. doi: 10.1007/BF00333412.
The relationship between the promoter length of the Kluyveromyces fragilis beta-glucosidase gene and the level of its expression in Saccharomyces cerevisiae was studied by gene fusion between deleted promoter fragments of various lengths and the promoterless beta-galactosidase gene of Escherichia coli. The removal of a region from position -425 to -232 led to a tenfold increase in the expression of the gene. The same results were obtained for the reconstructed beta-glucosidase gene with the same promoter length. It is likely that the deletion of this part of the promoter removes negative regulatory elements which are functional in Saccharomyces cerevisiae. This increase in activity is the main event which may explain the high increase in gene expression (60-fold) previously observed for an upstream deletion obtained during subcloning experiments of the beta-glucosidase gene. It is also shown that the expression of the gene greatly depends upon the nature of the recipient strain, the growth phase of the cell and that of the vector carrying it.
通过将不同长度的缺失启动子片段与大肠杆菌无启动子的β-半乳糖苷酶基因进行基因融合,研究了脆壁克鲁维酵母β-葡萄糖苷酶基因启动子长度与其在酿酒酵母中表达水平之间的关系。从-425位至-232位区域的缺失导致该基因表达增加了10倍。对于具有相同启动子长度的重构β-葡萄糖苷酶基因,也获得了相同的结果。很可能启动子这部分的缺失去除了在酿酒酵母中起作用的负调控元件。活性的这种增加是主要事件,这可能解释了先前在β-葡萄糖苷酶基因亚克隆实验中获得的上游缺失导致基因表达大幅增加(60倍)的现象。还表明,该基因的表达很大程度上取决于受体菌株的性质、细胞的生长阶段以及携带它的载体的生长阶段。