Sahu Satya Ranjan, Utkalaja Bhabasha Gyanadeep, Patel Shraddheya Kumar, Acharya Narottam
Laboratory of Genomic Instability and Diseases, Department of Infectious Disease Biology, Institute of Life Sciences, Bhubaneswar, India.
Bio Protoc. 2023 Nov 5;13(21):e4872. doi: 10.21769/BioProtoc.4872.
Cellular sensitivity is an approach to inhibit the growth of certain cells in response to any non-permissible conditions, as the presence of a cytotoxic agent or due to changes in growth parameters such as temperature, salt, or media components. Sensitivity tests are easy and informative assays to get insight into essential gene functions in various cellular processes. For example, cells having any functionally defective genes involved in DNA replication exhibit sensitivity to non-permissive temperatures and to chemical agents that block DNA replication fork movement. Here, we describe a sensitivity test for multiple strains of and of diverged genetic backgrounds subjected to several genotoxic chemicals simultaneously. We demonstrate it by testing the sensitivity of DNA polymerase defective yeast mutants by using spot analysis combined with colony forming unit (CFU) efficiency estimation. The method is very simple and inexpensive, does not require any sophisticated equipment, can be completed in 2-3 days, and provides both qualitative and quantitative data. We also recommend the use of this reliable methodology for assaying the sensitivity of these and other fungal species to antifungal drugs and xenobiotic factors.
细胞敏感性是一种在遇到任何不允许的条件时抑制某些细胞生长的方法，这些条件如细胞毒性剂的存在，或由于生长参数的变化，如温度、盐或培养基成分。敏感性测试是简单且信息丰富的分析方法，可深入了解各种细胞过程中的基本基因功能。例如，参与DNA复制的任何功能缺陷基因的细胞对非允许温度和阻断DNA复制叉移动的化学试剂表现出敏感性。在此，我们描述了一种针对具有不同遗传背景的多个菌株同时进行几种基因毒性化学物质处理的敏感性测试。我们通过使用斑点分析结合菌落形成单位（CFU）效率估计来测试DNA聚合酶缺陷型酵母突变体的敏感性来证明这一点。该方法非常简单且成本低廉，不需要任何复杂的设备，可在2至3天内完成，并提供定性和定量数据。我们还建议使用这种可靠的方法来测定这些和其他真菌物种对抗真菌药物和异源生物因子的敏感性。
