Tejos Ricardo, Osorio-Navarro Claudio, Norambuena Lorena
Facultad de Recursos Naturales Renovables, Universidad Arturo Prat, Iquique, Chile.
Department of Biology, Faculty of Sciences, Plant Molecular Biology Centre, Universidad de Chile, Santiago, Chile.
Methods Mol Biol. 2018;1789:143-154. doi: 10.1007/978-1-4939-7856-4_11.
Chemical compounds are useful to perturb biological functions in the same way as classical genetic approaches take advantage of mutations at the DNA level to perturb gene function. The use of bioactive chemicals currently called chemical genetic is especially valuable for cell biology. Chemical genetic approaches allow perturbations of cellular processes post-germination in a given time window controlling the severity of the effect by modifying or modulating the dose and/or the period of the treatment. Additionally, compounds can be applied directly to different mutants and translational fluorescent reporters/marker lines, expanding the repertoire of experimental setups addressing cell biology research. In this chapter, we describe standard protocols to visualize vacuole morphology and trafficking to the vacuole and the use of bioactive compounds as a proxy to study these biological processes.
化学化合物在干扰生物学功能方面很有用,其方式与经典遗传学方法利用DNA水平的突变来干扰基因功能相同。目前被称为化学遗传学的生物活性化学物质的使用对细胞生物学特别有价值。化学遗传学方法允许在给定的时间窗口内,在种子萌发后对细胞过程进行干扰,通过改变或调节剂量和/或处理时间来控制效应的严重程度。此外,化合物可以直接应用于不同的突变体和翻译荧光报告基因/标记系,扩展了用于细胞生物学研究的实验设置的范围。在本章中,我们描述了可视化液泡形态和向液泡运输的标准方案,以及使用生物活性化合物作为研究这些生物学过程的替代方法。