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中间碳代谢及碳分解代谢物阻遏解除缺陷的酵母突变体的分离与鉴定

Isolation and characterization of yeast mutants defective in intermediary carbon metabolism and in carbon catabolite derepression.

作者信息

Ciriacy M

出版信息

Mol Gen Genet. 1977 Jul 20;154(2):213-20. doi: 10.1007/BF00330840.

Abstract

Yeast mutants deficient in the constitutive ADHI (adc 1) were used for the isolation of mutants with deficiencies of the intermediary carbon metabolism, and of mutants defective in carbon catabolite derepression. Mutants were recognized by their inability to grow on YEP-glycerol and/or on ethanol synthetic complete medium. They were either defective in isocitrate lyase (ic11), succinate dehydrogenase (sdh1), or malate dehydrogenase (mdh1, mdh2), mdh-mutants could not uniformely be appointed to one of the known MDH isozymes. Homozygous mdh and sdh1 diploids are unable to sporulate. Three gene loci could be identified by mutants pleiotropically defective in many or all of the enzymes tested In ccr 1 mutants, derepression of isocitrate lyase, fructose-1,6-diphosphatase, ADHII and possibly of the cytoplasmic MDH is prevented, whereas the mitochondrial TCA-cycle enzymes, succinate dehydrogenase and malate dehydrogenase, are not significantly affected. CCR2 and CCR3 have quite similar action spectra. Both genes are obviously necessary for derepression of all enzymes tested. It could be shown that ccr1, ccr2 and ccr3 mutants are not respiratory deficient.

摘要

利用组成型乙醇脱氢酶1(adc1)缺陷的酵母突变体来分离中间碳代谢缺陷的突变体以及碳分解代谢物阻遏解除缺陷的突变体。通过它们在YEP-甘油和/或乙醇合成完全培养基上无法生长来识别突变体。它们要么异柠檬酸裂解酶(ic11)、琥珀酸脱氢酶(sdh1)有缺陷,要么苹果酸脱氢酶(mdh1、mdh2)有缺陷,mdh突变体不能一致地归为已知的MDH同工酶之一。纯合的mdh和sdh1二倍体无法形成孢子。在许多或所有测试酶中表现出多效性缺陷的突变体可鉴定出三个基因座。在ccr1突变体中,异柠檬酸裂解酶、果糖-1,6-二磷酸酶、乙醇脱氢酶II以及可能的细胞质MDH的阻遏解除受到阻碍,而线粒体三羧酸循环酶琥珀酸脱氢酶和苹果酸脱氢酶未受到显著影响。CCR2和CCR3具有相当相似的作用谱。这两个基因显然是所有测试酶阻遏解除所必需的。可以证明,ccr1、ccr2和ccr3突变体不存在呼吸缺陷。

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