Graf Katja, Hube Bernhard, Brunke Sascha
Department of Microbial Pathogenicity Mechanisms, Leibniz Institute for Natural Product Research and Infection Biology, Hans Knoell Institute, Jena, Germany.
Methods Mol Biol. 2021;2260:145-154. doi: 10.1007/978-1-0716-1182-1_10.
Experimental evolution is an experiment class of its own; instead of requiring an a priori hypothesis, the genetic adaptation of microbes to defined environments tells us about the underlying pathways and mechanisms. Such experiments are often deceptively simple in their design, based on a single abiotic stressor and what is in essence a long-term continuous culture. However, they generally provide a starting point to thorough follow-up analyses (which are specific for the organism at hand and not part of this method chapter). In this chapter, we describe a method to use a biotic stressor which is frequently encountered by pathogenic fungi-macrophage-like cells-in a serial passaging regime. Experimental evolution under such conditions can reveal new virulence attributes and mechanisms by selecting for adaptive mutations against the host cell-induced stress.It is important to note that every evolution experiment is different, and these techniques should be taken as a general guideline to be adapted to different organisms and questions. Then, it is a powerful tool with many potential applications in pathobiology research.
实验进化是一类独特的实验；微生物对特定环境的遗传适应并不需要先验假设，而是能告诉我们潜在的途径和机制。此类实验的设计往往看似简单，基于单一的非生物应激源以及本质上的长期连续培养。然而，它们通常为深入的后续分析提供了一个起点（这些分析针对手头的生物体，并非本章方法的一部分）。在本章中，我们描述一种在连续传代培养体系中使用病原真菌经常遇到的生物应激源——巨噬细胞样细胞的方法。在这种条件下的实验进化可以通过选择针对宿主细胞诱导应激的适应性突变来揭示新的毒力属性和机制。需要注意的是，每个进化实验都是不同的，这些技术应被视为一个通用指南，以适应不同的生物体和问题。因此，它是一种在病理生物学研究中有许多潜在应用的强大工具。
