Duplessis Martin, Russell W Michael, Romero Dennis A, Moineau Sylvain
Département de biochimie et de microbiologie, Faculté des sciences et de génie, Université Laval, Québec City, Canada.
Virology. 2005 Sep 30;340(2):192-208. doi: 10.1016/j.virol.2005.05.033.
A custom microarray was developed to study the temporal gene expression of the two groups of phages infecting the Gram-positive lactic acid bacterium Streptococcus thermophilus. The complete genomic sequence of the virulent cos-type phage DT1 (34,815 bp) and the pac-type phage 2972 (34,704 bp) were used for the construction of the microarray. Gene expression was measured at nine time intervals (0, 2, 7, 12, 17, 22, 27, 32 and 37 min) during phage infection and an expression curve was determined for each gene. Each phage gene was then classified into one of the three traditional transcription classes and these data were used to generate the complete transcriptional map of DT1 and 2972. Phage DT1 possesses 18 early genes, 12 middle genes and 12 late-expressed genes whereas 2972 has 16 early, 11 middle and 14 late genes. The trends of the phage gene expression profiles were also confirmed by slot blot hybridizations. Significant differences were observed when comparing the transcriptional maps of DT1 and 2972 with those already available for the S. thermophilus phages Sfi19 and Sfi21. To our knowledge, this report presents the first complete transcription analysis of bacteriophages infecting Gram-positive bacteria using the DNA microarray technology.
开发了一种定制微阵列，用于研究感染革兰氏阳性乳酸菌嗜热链球菌的两组噬菌体的基因表达时间变化。烈性cos型噬菌体DT1（34,815 bp）和pac型噬菌体2972（34,704 bp）的完整基因组序列用于构建微阵列。在噬菌体感染期间的九个时间间隔（0、2、7、12、17、22、27、32和37分钟）测量基因表达，并确定每个基因的表达曲线。然后将每个噬菌体基因分类为三个传统转录类别之一，并使用这些数据生成DT1和2972的完整转录图谱。噬菌体DT1拥有18个早期基因、12个中期基因和12个晚期表达基因，而2972有16个早期基因、11个中期基因和14个晚期基因。噬菌体基因表达谱的趋势也通过狭缝印迹杂交得到证实。将DT1和2972的转录图谱与嗜热链球菌噬菌体Sfi19和Sfi21的现有转录图谱进行比较时，观察到显著差异。据我们所知，本报告首次使用DNA微阵列技术对感染革兰氏阳性细菌的噬菌体进行了完整的转录分析。
