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通过分析mRNA水平研究乳酸克鲁维酵母中缺氧和氧化应激反应的方法。

An approach to the hypoxic and oxidative stress responses in Kluyveromyces lactis by analysis of mRNA levels.

作者信息

Blanco Moisés, Núñez Laura, Tarrío Nuria, Canto Elena, Becerra Manuel, González-Siso M Isabel, Cerdán M Esperanza

机构信息

Department of Molecular and Cell Biology, University of A Coruña, A Coruña, Spain.

出版信息

FEMS Yeast Res. 2007 Aug;7(5):702-14. doi: 10.1111/j.1567-1364.2007.00233.x. Epub 2007 Apr 10.

Abstract

Genome duplication, after the divergence of Saccharomyces cerevisiae from Kluyveromyces lactis along evolution, has been proposed as a mechanism of yeast evolution from strict aerobics, such as Candida albicans, to facultatives/fermentatives, such as S. cerevisiae. This feature, together with the preponderance of respiration and the use of the pentose phosphate pathway in glucose utilization, makes K. lactis a model yeast for studies related to carbon and oxygen metabolism. In this work, and based on the knowledge of the sequence of the genome of K. lactis, obtained by the Génolevures project, we have constructed DNA arrays from K. lactis including a limited amount of selected probes. They are related to the aerobiosis-hypoxia adaptation and to the oxidative stress response, and have been used to test changes in mRNA levels in response to hypoxia and oxidative stress generated by H(2)O(2). The study was carried out in both wild-type and rag2 mutant K. lactis strains in which glycolysis is blocked at the phosphoglucose isomerase step. This approach is the first analysis carried out in K. lactis for the majority of the genes selected.

摘要

在酿酒酵母与乳酸克鲁维酵母在进化过程中分化之后,基因组加倍被认为是酵母从严格需氧菌(如白色念珠菌)进化为兼性菌/发酵菌(如酿酒酵母)的一种机制。这一特性,连同呼吸作用的优势以及在葡萄糖利用中磷酸戊糖途径的使用,使得乳酸克鲁维酵母成为研究碳和氧代谢相关问题的模式酵母。在这项工作中,基于通过Génolevures项目获得的乳酸克鲁维酵母基因组序列知识,我们构建了包含有限数量选定探针的乳酸克鲁维酵母DNA阵列。它们与需氧-缺氧适应以及氧化应激反应相关,并已用于测试响应缺氧和H₂O₂产生的氧化应激时mRNA水平的变化。该研究在野生型和rag2突变型乳酸克鲁维酵母菌株中进行,其中糖酵解在磷酸葡萄糖异构酶步骤被阻断。对于大多数选定的基因而言,这种方法是在乳酸克鲁维酵母中进行的首次分析。

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