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[예방약학실험11] Ames test

"[예방약학실험11] Ames test"에 대한 내용입니다.
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최초등록일 2023.06.21 최종저작일 2023.06
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[예방약학실험11] Ames test
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  • 미리보기

    목차

    1. Experimental purpose

    2. Material
    A. Kit
    B. Apparatus
    C. Reagent

    3. Method

    4. Result

    5. Discussion
    A. TA100은 어떤 특성을 갖고 있는지 설명하시오.
    B. 만약 4번 sample에서 색 변화가 있었지만, 8번 sample에서는 색 변화가 일어나지 않았다면 compound K는 어떤 특성을 갖고 있는가?
    C. 결과를 보고 각자의 결론을 서술하세요.

    본문내용

    1. Experimental purpose: Ames test를 통해 compound K의 유전독성을 시험한다.
    2. Material
    A. Kit: Muta-ChromoPlate™ Kits (Growth media, 96 well plate, Ziploc bag, etc)
    B. Apparatus: lyophilized vial, incubator, pipet, tip, 1.5ml tube, tube rack, latex glove
    C. Reagent
    i. Strain: Salmonella (TA 100)
    ii. Control: NaN₃ (sodium azide)
    iii. Reaction mix: Davis-Mingoli Salt, D-glucose, Bromocresol purple, D-biotin, Lhistidine (극소량)

    참고자료

    · 없음

  • AI와 토픽 톺아보기

    • 1. Ames test
      The Ames test is a widely used bacterial reverse mutation assay that is considered a reliable and efficient method for evaluating the mutagenic potential of chemical compounds. It utilizes genetically modified strains of Salmonella typhimurium bacteria that are deficient in the ability to synthesize the amino acid histidine, making them unable to grow on histidine-deficient media. When exposed to a mutagenic compound, these bacteria can revert to a histidine-producing state, allowing them to grow on the selective media. The Ames test is valuable in assessing the genotoxicity of substances, as mutations in the bacterial genome can indicate the potential for a compound to cause genetic damage in higher organisms, including humans. The test is relatively simple, cost-effective, and can provide rapid results, making it a widely adopted tool in the field of toxicology and drug development. While it does not directly measure carcinogenicity, the Ames test serves as a useful screening method to identify potential mutagens and guide further, more comprehensive investigations.
    • 2. Salmonella typhimurium TA100
      Salmonella typhimurium TA100 is a widely used bacterial strain in the Ames test, a standard assay for evaluating the mutagenic potential of chemical compounds. This strain is genetically engineered to be deficient in the ability to synthesize the amino acid histidine, rendering it unable to grow on histidine-deficient media. When exposed to a mutagenic substance, the TA100 strain can undergo a reverse mutation, restoring its ability to produce histidine and allowing it to grow on the selective media. The TA100 strain is particularly sensitive to base-pair substitution mutations, making it a valuable tool for detecting a wide range of mutagenic compounds. Its use in the Ames test has contributed significantly to our understanding of chemical genotoxicity and has become a standard method in regulatory toxicology and drug development. The reliability and reproducibility of the TA100 strain have made it an essential component of the Ames test, providing a robust and well-characterized system for assessing the mutagenic potential of various substances.
    • 3. Compound K
      Compound K, also known as 20-O-β-D-glucopyranosyl-20(S)-protopanaxadiol, is a metabolite of the ginsenoside Rb1, which is a major active component of ginseng. Compound K has been the subject of extensive research due to its potential therapeutic properties, particularly in the areas of cancer, inflammation, and metabolic disorders. One of the key features of Compound K is its ability to exhibit anti-cancer effects through various mechanisms, such as inducing apoptosis, inhibiting cell proliferation, and suppressing angiogenesis in various cancer cell lines. Studies have shown that Compound K can effectively target and inhibit the growth of various types of cancer, including breast, prostate, colon, and lung cancer. Additionally, Compound K has demonstrated anti-inflammatory properties, which may be beneficial in the management of chronic inflammatory conditions. Furthermore, Compound K has been investigated for its potential in improving metabolic health. Some studies have suggested that Compound K may have the ability to enhance insulin sensitivity, regulate glucose homeostasis, and potentially alleviate symptoms associated with metabolic disorders, such as type 2 diabetes. Overall, Compound K is a promising natural compound with a wide range of therapeutic applications, and continued research in this area may lead to the development of novel treatments or adjuvant therapies for various health conditions.
    • 4. S9 fraction
      The S9 fraction is a crucial component in the Ames test and other in vitro genotoxicity assays. It is a metabolic activation system derived from the liver of rodents, typically rats or hamsters, that provides a source of metabolic enzymes to mimic the biotransformation of chemicals that may occur in the human body. The S9 fraction contains a mixture of cytochrome P450 enzymes, as well as other phase I and phase II metabolic enzymes, which can metabolize certain chemicals into more reactive or genotoxic metabolites. This is particularly important for the Ames test, as many compounds require metabolic activation to become mutagenic and induce reverse mutations in the bacterial strains used in the assay. The inclusion of the S9 fraction in the Ames test allows for the evaluation of both direct-acting mutagens and pro-mutagens, which require metabolic activation to exert their genotoxic effects. This is a significant advantage of the Ames test, as it provides a more comprehensive assessment of the mutagenic potential of chemical compounds. The use of the S9 fraction in the Ames test and other in vitro assays has been extensively validated and is considered a reliable approach to simulate the metabolic processes that may occur in the human body. It is an essential component in the assessment of chemical safety and the development of new drugs and other products.
    • 5. Bromocresol purple
      Bromocresol purple is a pH indicator dye that is commonly used in various analytical and biological applications. In the context of the Ames test, bromocresol purple plays a crucial role in the detection of bacterial growth and the identification of revertant colonies. In the Ames test, the bacterial strains used, such as Salmonella typhimurium TA100, are genetically modified to be histidine-dependent. When these bacteria are exposed to a mutagenic compound, they may undergo a reverse mutation, restoring their ability to synthesize histidine and allowing them to grow on histidine-deficient media. Bromocresol purple is added to the growth media in the Ames test to facilitate the visualization and enumeration of these revertant colonies. The dye changes color in response to changes in pH, with a purple color indicating a basic pH and a yellow color indicating an acidic pH. As the bacteria grow and metabolize the available nutrients, they produce acidic byproducts, causing a localized decrease in pH around the colonies. This pH change is detected by the bromocresol purple dye, which turns yellow, allowing the revertant colonies to be easily identified and counted. The use of bromocresol purple in the Ames test is a simple yet effective way to enhance the sensitivity and reliability of the assay. By providing a clear visual cue for the presence of revertant colonies, bromocresol purple contributes to the overall accuracy and reproducibility of the Ames test, which is a widely accepted and standardized method for evaluating the mutagenic potential of chemical compounds.

  • 자료후기

      Ai 리뷰
      Ames test를 통해 compound K의 돌연변이 유발 능력을 정확히 평가하였고, 실험 설계와 결과 분석이 체계적으로 이루어졌습니다.

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