Denis C L, Young E T
Mol Cell Biol. 1983 Mar;3(3):360-70. doi: 10.1128/mcb.3.3.360-370.1983.
The DNA segments containing the ADR1 gene and a mutant allele, ADR1-5c, have been isolated by complementation of function in Saccharomyces cerevisiae. The ADR1 gene is required for synthesis of the glucose-repressible alcohol dehydrogenase (ADHII) when S. cerevisiae cells are grown on a nonfermentable carbon source, whereas the ADR1-5c allele allows ADHII synthesis even during glucose repression. A plasmid pool consisting of yeast DNA fragments isolated from a strain carrying the ADR1-5c allele was used to transform a strain containing the adr1-1 allele, which prevents ADHII depression. Transformants were isolated which expressed ADHII during glucose repression. A plasmid isolated from one of these transformants was shown to carry the ADR1-5c allele by its ability to integrate at the chromosomal adr1-1 locus. The wild-type ADR1 gene was isolated by colony hybridization, using the cloned ADR1-5c gene as a probe. The ADR1-5c and ADR1 DNA segments were indistinguishable by restriction site mapping. A partial ADR1 phenotype could be conferred by a 1.9-kilobase region, but DNA outside of this region appeared to be necessary for normal activation of ADHII by the ADR1 gene.
通过在酿酒酵母中进行功能互补，已分离出包含ADR1基因和突变等位基因ADR1 - 5c的DNA片段。当酿酒酵母细胞在非发酵碳源上生长时，ADR1基因是合成葡萄糖可抑制的乙醇脱氢酶（ADHII）所必需的，而ADR1 - 5c等位基因即使在葡萄糖抑制期间也允许ADHII的合成。由从携带ADR1 - 5c等位基因的菌株中分离的酵母DNA片段组成的质粒库被用于转化含有adr1 - 1等位基因的菌株，该等位基因可防止ADHII的抑制。分离出了在葡萄糖抑制期间表达ADHII的转化体。从这些转化体之一中分离出的一个质粒通过其整合到染色体adr1 - 1位点的能力被证明携带ADR1 - 5c等位基因。使用克隆的ADR1 - 5c基因作为探针，通过菌落杂交分离出野生型ADR1基因。通过限制性酶切位点图谱分析，ADR1 - 5c和ADR1 DNA片段无法区分。一个1.9千碱基的区域可赋予部分ADR1表型，但该区域之外的DNA似乎是ADR1基因正常激活ADHII所必需的。
