β-半乳糖苷酶活性的菌落升降比色测定法

IF 2.7 3区环境科学与生态学 Q3 ENVIRONMENTAL SCIENCES

Journal of Arid Land Pub Date : 2016-12-01 DOI:10.1101/pdb.prot088963

Juan I Fuxman Bass, John S Reece-Hoyes, Albertha J M Walhout

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引用次数: 0

摘要

在本方案中，我们介绍了一种定性检测方法，用于监测 LacZ 基因编码的 β-半乳糖苷酶在酵母中的表达水平。这种方法既可用于确定酵母 DNA "诱饵 "菌株中 LacZ 表达的自身活性，也可用于评估酵母单杂交（Y1H）试验中转录因子 "猎物 "与感兴趣的 DNA 诱饵相互作用所介导的 LacZ 报告基因激活。在这种比色测定法中，酵母在液氮中裂解，然后用无色化合物 X-gal（在这种酶的存在下会变成蓝色）测定β-半乳糖苷酶的表达。

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Colony Lift Colorimetric Assay for β-Galactosidase Activity.

In this protocol, we present a qualitative assay for monitoring the level of expression of β-galactosidase, an enzyme encoded by the LacZ gene, in yeast. This is useful both for determining autoactivity of LacZ expression in yeast DNA "bait" strains and for assessing LacZ reporter gene activation mediated by a transcription factor "prey" interaction with a DNA bait of interest in yeast one-hybrid (Y1H) assays. In this colorimetric assay, yeast are lysed in liquid nitrogen and then assayed for β-galactosidase expression using the colorless compound X-gal, which turns blue in the presence of this enzyme.

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来源期刊

Journal of Arid Land ENVIRONMENTAL SCIENCES-

CiteScore

4.70

自引率

6.70%

发文量

768

审稿时长

3.2 months

期刊介绍： The Journal of Arid Land is an international peer-reviewed journal co-sponsored by Xinjiang Institute of Ecology and Geography, the Chinese Academy of Sciences and Science Press. It aims to meet the needs of researchers, students and practitioners in sustainable development and eco-environmental management, focusing on the arid and semi-arid lands in Central Asia and the world at large. The Journal covers such topics as the dynamics of natural resources (including water, soil and land, organism and climate), the security and sustainable development of natural resources, and the environment and the ecology in arid and semi-arid lands, especially in Central Asia. Coverage also includes interactions between the atmosphere, hydrosphere, biosphere, and lithosphere, and the relationship between these natural processes and human activities. Also discussed are patterns of geography, ecology and environment; ecological improvement and environmental protection; and regional responses and feedback mechanisms to global change. The Journal of Arid Land also presents reviews, brief communications, trends and book reviews of work on these topics.