Chen Feng, Wang Kui, Stewart Jeneen, Belas Robert
Center of Marine Biotechnology, University of Maryland Biotechnology Institute, Baltimore, MD 21202, USA.
Appl Environ Microbiol. 2006 Jul;72(7):4995-5001. doi: 10.1128/AEM.00056-06.
Approximately 70% of sequenced bacterial genomes contain prophage-like structures, yet little effort has been made to use this information to determine the functions of these elements. The recent genomic sequencing of the marine bacterium Silicibacter sp. strain TM1040 revealed five prophage-like elements in its genome. The genomes of these prophages (named prophages 1 to 5) are approximately 74, 30, 39, 36, and 15 kb long, respectively. To understand the function of these prophages, cultures of TM1040 were treated with mitomycin C to induce the production of viral particles. A significant increase in viral counts and a decrease in bacterial counts when treated with mitomycin C suggested that prophages were induced from TM1040. Transmission electron microscopy revealed one dominant type of siphovirus, while pulsed-field gel electrophoresis demonstrated two major DNA bands, equivalent to 35 and 75 kb, in the lysate. PCR amplification with primer sets specific to each prophage detected the presence of prophages 1, 3, and 4 in the viral lysate, suggesting that these prophages are inducible, but not necessarily to the same level, while prophages 2 and 5 are likely defective or non-mitomycin C-inducible phages. The combination of traditional phage assays and modern microbial genomics provides a quick and efficient way to investigate the functions and inducibility of prophages, particularly for a host harboring multiple prophages with similar sizes and morphological features.
大约70%已测序的细菌基因组含有类前噬菌体结构,但利用这些信息来确定这些元件的功能的工作却做得很少。最近对海洋细菌硅细菌属菌株TM1040进行的基因组测序揭示了其基因组中有五个类前噬菌体元件。这些前噬菌体(命名为前噬菌体1至5)的基因组长度分别约为74、30、39、36和15 kb。为了了解这些前噬菌体的功能,用丝裂霉素C处理TM1040的培养物以诱导病毒颗粒的产生。用丝裂霉素C处理后病毒数量显著增加而细菌数量减少,这表明前噬菌体是从TM1040中诱导产生的。透射电子显微镜显示一种主要类型的肌尾噬菌体,而脉冲场凝胶电泳在裂解物中显示出两条主要的DNA条带,相当于35和75 kb。用针对每个前噬菌体的引物组进行PCR扩增,在病毒裂解物中检测到前噬菌体1、3和4的存在,这表明这些前噬菌体是可诱导的,但诱导水平不一定相同,而前噬菌体2和5可能是缺陷型或非丝裂霉素C诱导型噬菌体。传统噬菌体检测方法与现代微生物基因组学相结合,为研究前噬菌体的功能和诱导性提供了一种快速有效的方法,特别是对于含有多个大小和形态特征相似的前噬菌体的宿主。