High permeability bread yeast for detecting environmental carcinogen and preparation method thereof

A technology for baker's yeast and detection environment, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganism measurement/inspection, etc., can solve the problems of baker's yeast cell wall permeability barrier, lower system detection sensitivity, detection , to achieve the effects of monitoring, improving detection sensitivity, and expanding the detection range

Inactive Publication Date: 2008-08-13
INST OF AQUATIC LIFE ACAD SINICA
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Problems solved by technology

Genotoxic carcinogens in the environment usually exist in very low concentrations. Due to the permeability barrier of baker's yeast cell wall, the accumulation of environmental genotoxic carcinogens in baker's yeast cells is limited, thereby reducing the detection sensitivity of the system; large molecular weight The chemical does not even pass freely through the baker's yeast cell wall and cannot be detected by the system

Method used

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  • High permeability bread yeast for detecting environmental carcinogen and preparation method thereof

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Embodiment Construction

[0017] The high-permeability baker's yeast that can be used to detect environmental carcinogens of the present invention is characterized in that the high-permeability baker's yeast is a baker's yeast that lacks CWP1 and CWP2 double genes.

[0018] The specific steps for preparing highly permeable baker's yeast that can be used to detect environmental carcinogens are as follows:

[0019] 1. Extraction of baker's yeast genomic DNA

[0020] 1a. Take a small amount of baker's yeast (Saccharomyces.cerevisiae, haploid BY4741, MATa his3Δ1leu2Δ0met15Δ0ura3Δ0), inoculate it in 5 ml of YPD culture medium, culture it at 30°C and 200 rpm for 16 hours, and obtain a baker's yeast liquid;

[0021] Wherein: the formula of YPD culture medium is: yeast extract 1%, peptone 2%, glucose 2%, and the rest is water;

[0022] 1b. Take 1.5 ml of baker's yeast liquid, centrifuge at 4000 rpm for 2 minutes, and suspend the precipitate in 200 microliters of extraction buffer;

[0023] Wherein: the formu...

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Abstract

The invention discloses a high permeability baker's yeast for detecting environmental carcinogen and method for making the same which relates to baker's yeast and the reforming of descendiblity gene. The permeability of the baker's yeast used for detecting environmental carcinogen is improved by the reforming of descendiblity gene so that the baker's yeast can detect the environmental carcinogen in more large scale and in higher sensitivity. The gene CWP1 and CWP2 of encoded baker's yeast cell wall important structural protein mannose egg albumen are inactivated by gene out-knocking technology to improve the permeability of baker's yeast cell for environmental carcinogen because the baker's yeast cell wall is the main disorder for chemistry permeability. A RNR3-lacZ system is constructed in CWP1 and CWP2 gene out-knocking baker's yeast saccharomycete of reformed baker's yeast cell wall to improve the sensitivity for detecting environment genotoxic carcinogens.

Description

Technical field: [0001] The invention relates to baker's yeast. The baker's yeast, which can be used to detect environmental carcinogens, is genetically modified to improve the permeability, so that the detection range of environmental carcinogens is wider and the sensitivity is higher. Background technique: [0002] Harmful industrial chemicals in the environment are one of the causes of cancer. These harmful industrial chemicals can damage DNA, induce DNA mutations, and cause cancer. This kind of chemical that can react with the genetic material of cells, can directly or through metabolic activation to generate electrophilic substances, change the structure and function of DNA, and induce tumors by proliferation is called genotoxic carcinogen. These carcinogens have obvious mutagenic effects, can cause cell mutation or chromosomal damage, and are more harmful. Therefore, monitoring these genotoxic carcinogens in the environment is of great significance for controlling and...

Claims

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Application Information

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IPC IPC(8): C12N1/19C12N15/52C12N15/81C12Q1/04C12Q1/34C12R1/645
Inventor 肖伟戴和平张敏张晓华
Owner INST OF AQUATIC LIFE ACAD SINICA
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