Shan Jinyu, Clokie Martha
Department of Biological Sciences, University of Warwick, Coventry, UK.
Methods Mol Biol. 2009;502:171-6. doi: 10.1007/978-1-60327-565-1_10.
Bacteriophages manipulate bacterial gene expression in order to express their own genes or influence bacterial metabolism. Gene expression can be studied using real-time PCR or microarrays. Either technique requires the prior isolation of high quality RNA uncontaminated by the presence of genomic DNA. We outline the considerations necessary when working with bacteriophage infected bacterial cells. We also give an example of a protocol for extraction and quantification of high quality RNA from infected bacterial cells, using the marine cyanobacterium WH7803 and the phage S-PM2 as a case study. This protocol can be modified to extract RNA from the host/bacteriophage of interest.
噬菌体通过操纵细菌基因表达来表达自身基因或影响细菌代谢。基因表达可使用实时聚合酶链反应(PCR)或微阵列进行研究。这两种技术都需要事先分离出未被基因组DNA污染的高质量RNA。我们概述了处理噬菌体感染的细菌细胞时所需考虑的因素。我们还给出了一个使用海洋蓝细菌WH7803和噬菌体S-PM2作为案例研究,从感染的细菌细胞中提取和定量高质量RNA的方案示例。该方案可进行修改,以从感兴趣的宿主/噬菌体中提取RNA。