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利用酿酒酵母中的亚硫酸盐抗性作为显性选择标记。

Use of sulfite resistance in Saccharomyces cerevisiae as a dominant selectable marker.

作者信息

Park H, Lopez N I, Bakalinsky A T

机构信息

Department of Food Science and Technology, Oregon State University, Corvallis 97331-6602, USA.

出版信息

Curr Genet. 1999 Dec;36(6):339-44. doi: 10.1007/s002940050508.

Abstract

Two S. cerevisiae genes were found to exhibit dominant phenotypes useful for selecting transformants of industrial and laboratory strains of S. cerevisiae. FZF1-4, which confers sulfite resistance, was originally isolated and identified as RSU1-4, but the two genes are shown here to be allelic. Cysteine 57 in wild-type Fzf1p was found to be replaced by tyrosine in Fzf1-4p. Multicopy SSU1, which also confers sulfite resistance, was found to be somewhat less efficient. In both cases, a period of outgrowth in non-selective medium following transformation was found to be necessary. The number of transformants obtained was found to be strain-dependent, and also to depend on the sulfite concentration used during selection. Undesirable background growth of non-transformants was not observed at cell densities as high as 2.5 x 10(7)/plate. In two ura3 laboratory strains where selection for URA3 was applied independently of that for sulfite, the transformation efficiency for sulfite resistance was about 50% that for uracil prototrophy.

摘要

发现两个酿酒酵母基因表现出显性表型,有助于筛选酿酒酵母工业菌株和实验室菌株的转化体。赋予亚硫酸盐抗性的FZF1 - 4最初被分离并鉴定为RSU1 - 4,但本文表明这两个基因是等位基因。发现野生型Fzf1p中的半胱氨酸57在Fzf1 - 4p中被酪氨酸取代。同样赋予亚硫酸盐抗性的多拷贝SSU1效率略低。在这两种情况下,发现转化后在非选择性培养基中生长一段时间是必要的。获得的转化体数量取决于菌株,也取决于选择过程中使用的亚硫酸盐浓度。在细胞密度高达2.5×10(7)/平板时,未观察到非转化体的不良背景生长。在两个独立于亚硫酸盐选择进行URA3选择的ura3实验室菌株中,亚硫酸盐抗性的转化效率约为尿嘧啶原养型转化效率的50%。

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