Gay-Andrieu Françoise, Alex Deepu, Calderone Richard
Department of Pathology, GU-MedStar Hospital, Georgetown University Medical Center, Washington DC, USA.
Department of Microbiology & Immunology, Georgetown University Medical Center, Washington DC, 20057, USA.
Methods Mol Biol. 2016;1356:165-72. doi: 10.1007/978-1-4939-3052-4_12.
The search for new antifungal drugs and cell targets continues. During the discovery process, mechanism-of-action (MOA) studies are critical to the continued progress of the compound through the pipeline. There are many approaches that can be utilized in understanding the MOA. One of these approaches is a genetic screen utilizing the availability of Saccharomyces cerevisiae mutant libraries. Both null and heterozygous library mutants covering the entire genome of this model yeast are available. The desired phenotype when screening the new compound is either resistance (null mutants) or haploinsufficiency or loss of fitness (heterozygote mutants). Both types of mutants can be clustered by software into common targets that provide clues as to a pathway or other cell process. Below, methods are described for genetic screens.
对新型抗真菌药物和细胞靶点的研究仍在继续。在发现过程中，作用机制（MOA）研究对于化合物在研发流程中的持续推进至关重要。有许多方法可用于理解作用机制。其中一种方法是利用酿酒酵母突变体文库进行遗传筛选。覆盖这种模式酵母整个基因组的纯合和杂合文库突变体均有可得。筛选新化合物时所需的表型要么是抗性（纯合突变体），要么是单倍体不足或适应性丧失（杂合子突变体）。这两种类型的突变体都可以通过软件聚类为共同靶点，从而为某一途径或其他细胞过程提供线索。以下将介绍遗传筛选的方法。
