Gilbert Jack A, Hughes Margaret
Plymouth Marine Laboratory, The Hoe, Plymouth, UK.
Methods Mol Biol. 2011;733:195-205. doi: 10.1007/978-1-61779-089-8_14.
Metatranscriptomics has been developed to help understand how communities respond to changes in their environment. Metagenomic studies provided a snapshot of the genetic composition of the community at any given time. However, short-timescale studies investigating the response of communities to rapid environmental changes (e.g. pollution events or diurnal light availability) require analysis of changes in the abundance and composition of the active fraction of the community. Metatranscriptomics enables researchers to investigate the actively transcribed ribosomal and messenger RNA from a community. It has been applied to environments as diverse as soil and seawater. This chapter outlines sampling protocols and RNA extraction techniques from these two ecosystems, as well as details a method to enrich mRNA in the extracted nucleic acid. Also, a section is dedicated for outlining a bioinformatic procedure for the analysis of metatranscriptomic datasets.
宏转录组学已得到发展,以帮助了解群落如何对其环境变化做出反应。宏基因组学研究提供了任何给定时间群落遗传组成的快照。然而,研究群落对快速环境变化(如污染事件或昼夜光照可用性)反应的短期研究需要分析群落活性部分的丰度和组成变化。宏转录组学使研究人员能够研究群落中活跃转录的核糖体RNA和信使RNA。它已应用于土壤和海水等多种环境。本章概述了从这两个生态系统中采样的方案和RNA提取技术,并详细介绍了一种在提取的核酸中富集mRNA的方法。此外,还有一节专门概述了用于分析宏转录组学数据集的生物信息学程序。