Kawai S, Hikiji T, Murao S, Takagi M, Yano K
Department of Agricultural Chemistry, University of Tokyo, Japan.
Agric Biol Chem. 1991 Jan;55(1):59-65.
The host-vector systems of an n-alkane-assimilating-yeast, Candida maltosa, that we previously constructed consisted of a vector replicating with an ARS region of this yeast, and C. maltosa strains J288 (leu2) or CH1 (his5) as hosts. Since each of these hosts has a single genetic marker, we have developed a new host-vector system using two genetic markers. By UV irradiation of strain CH1, an adenine auxotrophic mutant, CHA1, forming red colonies was isolated. A DNA fragment complementing this deficiency was isolated from the C. maltosa genome. Since the DNA fragment also complemented the ade1 mutation of S. cerevisiae, we termed a gene contained in this DNA fragment C-ADE1. The nucleotides of C-ADE1 were sequenced. The deduced amino acid sequence (291 residues) had 65.6% homology with that of ADE1 of S. cerevisiae (306 residues). Having the cloned C-ADE1 DNA, we improved the host-vector system of C. maltosa.
我们之前构建的一种正构烷烃同化酵母——麦芽糖假丝酵母的宿主-载体系统，由一个能与该酵母的自主复制序列（ARS）区域一起复制的载体，以及作为宿主的麦芽糖假丝酵母菌株J288（亮氨酸2缺陷型）或CH1（组氨酸5缺陷型）组成。由于这些宿主各自都有一个单一的遗传标记，我们利用两个遗传标记开发了一种新的宿主-载体系统。通过对CH1菌株进行紫外线照射，分离出了一个形成红色菌落的腺嘌呤营养缺陷型突变体CHA1。从麦芽糖假丝酵母基因组中分离出了一个能弥补这种缺陷的DNA片段。由于该DNA片段也能弥补酿酒酵母的ade1突变，我们将这个DNA片段中包含的基因命名为C-ADE1。对C-ADE1的核苷酸进行了测序。推导的氨基酸序列（291个残基）与酿酒酵母ADE1的氨基酸序列（306个残基）有65.6%的同源性。拥有克隆的C-ADE1 DNA后，我们改进了麦芽糖假丝酵母的宿主-载体系统。
