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酿酒酵母Ca2+敏感突变体的分离与鉴定

Isolation and characterization of Ca2+-sensitive mutants of Saccharomyces cerevisiae.

作者信息

Ohya Y, Ohsumi Y, Anraku Y

出版信息

J Gen Microbiol. 1986 Apr;132(4):979-88. doi: 10.1099/00221287-132-4-979.

Abstract

Thirty Ca2+-sensitive (cls: calcium sensitive) mutants of Saccharomyces cerevisiae were isolated by replica-plating. These mutants, which each had a single recessive chromosomal mutation, were divided into 18 complementation groups. Some cls mutants showed a phenotype of specific sensitivity to Ca2+, while others showed phenotypes of sensitivities to several divalent cations. From measurements of the calcium contents and initial rates of Ca2+ uptake of the cls mutants, 16 of the 18 cls complementation groups were classified into four types: type I mutants (cls5, cls6, cls13, cls14, cls15, cls16, cls17, and cls18) had both elevated calcium contents and increased uptake activities. A type II mutant (cls4) had a normal calcium content and normal uptake activity; type III mutants (cls1, cls2 and cls3) had elevated calcium contents but normal initial rates of Ca2+ uptake; type IV mutants (cls8, cls9, cls10 and cls11) had normal calcium contents but increased initial rates of Ca2+ uptake. Two of the mutants (cls7 and cls12) had intermediate biochemical properties. The primary defects of these four types of cls mutants were considered in terms of the Ca2+ transport system(s). Both type I and type III mutants, which had elevated calcium contents, simultaneously showed a trifluoperazine-sensitive phenotype, suggesting a close correlation of this phenotype with elevated calcium content. In addition, all type IV mutants were unable to utilize nonfermentable sugars. One CLS gene, CLS7, was located on the left arm of chromosome V.

摘要

通过影印平板法分离出了30个酿酒酵母的钙离子敏感(cls:钙敏感)突变体。这些突变体各自都有一个单一的隐性染色体突变,被分为18个互补群。一些cls突变体表现出对Ca2+的特异性敏感表型,而其他一些则表现出对几种二价阳离子敏感的表型。通过对cls突变体的钙含量和Ca2+摄取初始速率的测量,18个cls互补群中的16个被分为四种类型:I型突变体(cls5、cls6、cls13、cls14、cls15、cls16、cls17和cls18)的钙含量升高且摄取活性增加。II型突变体(cls4)的钙含量正常且摄取活性正常;III型突变体(cls1、cls2和cls3)的钙含量升高但Ca2+摄取初始速率正常;IV型突变体(cls8、cls9、cls10和cls11)的钙含量正常但Ca2+摄取初始速率增加。其中两个突变体(cls7和cls12)具有中间生化特性。从Ca2+转运系统的角度考虑了这四种类型cls突变体的主要缺陷。I型和III型突变体的钙含量都升高,同时表现出对三氟拉嗪敏感的表型,表明这种表型与钙含量升高密切相关。此外,所有IV型突变体都不能利用非发酵糖。一个CLS基因,CLS7,位于第五条染色体的左臂上。

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