Borglin Sharon, Joyner Dominique, Jacobsen Janet, Mukhopadhyay Aindrila, Hazen Terry C
Ecology Department, Earth Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
J Microbiol Methods. 2009 Feb;76(2):159-68. doi: 10.1016/j.mimet.2008.10.003. Epub 2008 Oct 22.
Growing anaerobic microorganisms in phenotypic microarrays (PM) and 96-well microtiter plates is an emerging technology that allows high throughput survey of the growth and physiology and/or phenotype of cultivable microorganisms. For non-model bacteria, a method for phenotypic analysis is invaluable, not only to serve as a starting point for further evaluation, but also to provide a broad understanding of the physiology of an uncharacterized wild-type organism or the physiology/phenotype of a newly created mutant of that organism. Given recent advances in genetic characterization and targeted mutations to elucidate genetic networks and metabolic pathways, high-throughput methods for determining phenotypic differences are essential. Here we outline challenges presented in studying the physiology and phenotype of a sulfate-reducing anaerobic delta proteobacterium, Desulfovibrio vulgaris Hildenborough. Modifications of the commercially available OmniLog system (Hayward, CA) for experimental setup, and configuration, as well as considerations in PM data analysis are presented. Also highlighted here is data viewing software that enables users to view and compare multiple PM data sets. The PM method promises to be a valuable strategy in our systems biology approach to D. vulgaris studies and is readily applicable to other anaerobic and aerobic bacteria.
在表型微阵列(PM)和96孔微量滴定板中培养厌氧微生物是一项新兴技术,它能够对可培养微生物的生长、生理学特性和/或表型进行高通量研究。对于非模式细菌而言,一种表型分析方法不仅是进一步评估的起点,而且对于全面了解未表征野生型生物体的生理学特性或该生物体新创建突变体的生理学特性/表型都具有重要价值。鉴于在基因表征和靶向突变以阐明遗传网络和代谢途径方面的最新进展,用于确定表型差异的高通量方法至关重要。在此,我们概述了在研究硫酸盐还原厌氧δ变形菌——希氏脱硫弧菌(Desulfovibrio vulgaris Hildenborough)的生理学特性和表型时所面临的挑战。介绍了针对市售OmniLog系统(加利福尼亚州海沃德)在实验设置、配置方面的改进,以及PM数据分析中的注意事项。这里还重点介绍了一种数据查看软件,该软件能让用户查看和比较多个PM数据集。PM方法有望成为我们研究希氏脱硫弧菌的系统生物学方法中的一种有价值策略,并且很容易应用于其他厌氧和好氧细菌。