Jackson R W, Giddens S R
Department of Biology and Biochemistry, University of Bath, Claverton Down, Bath, BA2 7AY, UK. RJ218@ bath.ac.uk
Infect Disord Drug Targets. 2006 Sep;6(3):207-40. doi: 10.2174/187152606778249944.
Establishing the mechanisms by which microbes interact with their environment, including eukaryotic hosts, is a major challenge that is essential for the economic utilisation of microbes and their products. Techniques for determining global gene expression profiles of microbes, such as microarray analyses, are often hampered by methodological restraints, particularly the recovery of bacterial transcripts (RNA) from complex mixtures and rapid degradation of RNA. A pioneering technology that avoids this problem is In Vivo Expression Technology (IVET). IVET is a 'promoter-trapping' methodology that can be used to capture nearly all bacterial promoters (genes) upregulated during a microbe-environment interaction. IVET is especially useful because there is virtually no limit to the type of environment used (examples to date include soil, oomycete, a host plant or animal) to select for active microbial promoters. Furthermore, IVET provides a powerful method to identify genes that are often overlooked during genomic annotation, and has proven to be a flexible technology that can provide even more information than identification of gene expression profiles. A derivative of IVET, termed resolvase-IVET (RIVET), can be used to provide spatio-temporal information about environment-specific gene expression. More recently, niche-specific genes captured during an IVET screen have been exploited to identify the regulatory mechanisms controlling their expression. Overall, IVET and its various spin-offs have proven to be a valuable and robust set of tools for analysing microbial gene expression in complex environments and providing new targets for biotechnological development.
确定微生物与包括真核宿主在内的环境相互作用的机制是一项重大挑战，这对于微生物及其产物的经济利用至关重要。诸如微阵列分析等用于确定微生物全球基因表达谱的技术，常常受到方法学限制的阻碍，特别是从复杂混合物中回收细菌转录本（RNA）以及RNA的快速降解。一种避免此问题的开创性技术是体内表达技术（IVET）。IVET是一种“启动子捕获”方法，可用于捕获在微生物与环境相互作用期间上调的几乎所有细菌启动子（基因）。IVET特别有用，因为用于选择活性微生物启动子的环境类型几乎没有限制（迄今为止的例子包括土壤、卵菌、宿主植物或动物）。此外，IVET提供了一种强大的方法来鉴定在基因组注释过程中经常被忽视的基因，并且已被证明是一种灵活的技术，它能提供比鉴定基因表达谱更多的信息。IVET的一种衍生物，称为解离酶-IVET（RIVET），可用于提供有关环境特异性基因表达的时空信息。最近，在IVET筛选过程中捕获的特定生态位基因已被用于鉴定控制其表达的调控机制。总体而言，IVET及其各种衍生技术已被证明是一套有价值且强大的工具，用于分析复杂环境中的微生物基因表达，并为生物技术发展提供新的靶点。
