Klein R D, Roof L L
Molecular Biology Research, Upjohn Company, Kalamazoo, MI 49007.
Curr Genet. 1988;13(1):29-35. doi: 10.1007/BF00365753.
A DNA fragment containing the gene encoding orotidine 5'-phosphate decarboxylase (ODC) from the yeast, Schwanniomyces occidentalis (formerly castellii) has been isolated from a genomic library constructed in the S. cerevisiae expression vector, pYcDE8. A recombinant plasmid, p2-1A, containing a 2.47 kb insert was shown to complement the ura3-52 mutation of several strains of S. cerevisiae. This DNA insert was shown to be from Schwanniomyces occidentalis by Southern hybridization analysis. A restriction enzyme cleavage map of the insert has been derived and the ODC gene localized to a 1.1 kb region by deletion analysis. In addition, we have demonstrated that expression of ODC is not dependent on the ADH1 promoter carried on pYcDE8. This is the first report of the cloning of a gene from a member of the genus Schwanniomyces.
从构建于酿酒酵母表达载体pYcDE8中的基因组文库中分离出了一个包含来自西方许旺酵母（原名为卡氏酵母）的编码乳清苷5'-磷酸脱羧酶（ODC）基因的DNA片段。一个含有2.47 kb插入片段的重组质粒p2-1A，被证明能够互补几株酿酒酵母的ura3-52突变。通过Southern杂交分析表明该DNA插入片段来自西方许旺酵母。已得出该插入片段的限制性酶切图谱，并通过缺失分析将ODC基因定位到一个1.1 kb的区域。此外，我们还证明了ODC的表达不依赖于pYcDE8上携带的ADH1启动子。这是关于从许旺酵母属成员中克隆基因的首次报道。
