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一种改良的麦芽糖假丝酵母宿主-载体系统,其使用从该酵母基因组中分离出的一个基因,该基因可互补酿酒酵母的his5突变。

An improved host-vector system for Candida maltosa using a gene isolated from its genome that complements the his5 mutation of Saccharomyces cerevisiae.

作者信息

Hikiji T, Ohkuma M, Takagi M, Yano K

机构信息

Department of Agricultural Chemistry, University of Tokyo, Japan.

出版信息

Curr Genet. 1989 Oct;16(4):261-6. doi: 10.1007/BF00422112.

Abstract

The host-vector system of an n-alkane-assimilating-yeast, Candida maltosa, which we previously constructed using an autonomously replicating sequence (ARS) region isolated from the genome of this yeast, utilizes C. maltosa J288 (leu2-) as a host. As this host had a serious growth defect on n-alkane, we developed an improved host-vector system using C. maltosa CH1 (his-) as host. The vectors were constructed with the Candida ARS region and a DNA fragment isolated from the genome of C. maltosa. Since this DNA fragment could complement histidine auxotrophy of both C. maltosa CH1 and S. cerevisiae (his5-), we termed the gene contained in this DNA fragment C-HIS5. The vectors were characterized in terms of transformation frequency and stability, and the nucleotide sequence of C-HIS5 was determined. The deduced amino acid sequence (389 residues) shared 51% homology with that of HIS5 of S. cerevisiae (384 residues; Nishiwaki et al. 1987).

摘要

我们之前利用从解脂假丝酵母基因组中分离出的自主复制序列(ARS)区域构建了一种解脂假丝酵母(Candida maltosa)的宿主-载体系统,该系统使用解脂假丝酵母J288(leu2-)作为宿主。由于该宿主在正构烷烃上存在严重的生长缺陷,我们开发了一种以解脂假丝酵母CH1(his-)作为宿主的改良宿主-载体系统。这些载体是用解脂假丝酵母的ARS区域和从解脂假丝酵母基因组中分离出的一个DNA片段构建的。由于这个DNA片段可以互补解脂假丝酵母CH1和酿酒酵母(his5-)的组氨酸营养缺陷型,我们将这个DNA片段中包含的基因命名为C-HIS5。对这些载体的转化频率和稳定性进行了表征,并测定了C-HIS5的核苷酸序列。推导的氨基酸序列(389个残基)与酿酒酵母HIS5的氨基酸序列(384个残基;Nishiwaki等人,1987年)有51%的同源性。

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