University of Würzburg, Biocentre, Department of Microbiology, Am Hubland, D-97074 Würzburg, Germany.
Nucleic Acids Res. 2010 Jan;38(3):868-77. doi: 10.1093/nar/gkp1032. Epub 2009 Nov 18.
Chlamydia trachomatis is an obligate intracellular pathogenic bacterium that has been refractory to genetic manipulations. Although the genomes of several strains have been sequenced, very little information is available on the gene structure of these bacteria. We used deep sequencing to define the transcriptome of purified elementary bodies (EB) and reticulate bodies (RB) of C. trachomatis L2b, respectively. Using an RNA-seq approach, we have mapped 363 transcriptional start sites (TSS) of annotated genes. Semi-quantitative analysis of mapped cDNA reads revealed differences in the RNA levels of 84 genes isolated from EB and RB, respectively. We have identified and in part confirmed 42 genome- and 1 plasmid-derived novel non-coding RNAs. The genome encoded non-coding RNA, ctrR0332 was one of the most abundantly and differentially expressed RNA in EB and RB, implying an important role in the developmental cycle of C. trachomatis. The detailed map of TSS in a thus far unprecedented resolution as a complement to the genome sequence will help to understand the organization, control and function of genes of this important pathogen.
沙眼衣原体是一种专性细胞内寄生的细菌，其遗传操作一直难以实现。尽管已经对多个菌株的基因组进行了测序，但对于这些细菌的基因结构却知之甚少。我们分别使用深度测序来定义纯化的原体（EB）和网状体（RB）的沙眼衣原体 L2b 的转录组。通过 RNA-seq 方法，我们共定位了 363 个注释基因的转录起始位点（TSS）。对映射的 cDNA 读数进行半定量分析，分别揭示了来自 EB 和 RB 的 84 个基因的 RNA 水平存在差异。我们已经鉴定并部分确认了 42 个源自基因组和质粒的新型非编码 RNA。基因组编码的非编码 RNA ctrR0332 在 EB 和 RB 中是表达丰度最高和差异表达最大的 RNA 之一，这暗示了它在沙眼衣原体发育周期中具有重要作用。这种空前分辨率的 TSS 详细图谱作为基因组序列的补充，将有助于理解该重要病原体基因的组织、调控和功能。
