Deep sequencing-based discovery of the Chlamydia trachomatis transcriptome.

Nucleic Acids Research
Marco AlbrechtThomas Rudel

Abstract

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.

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Methods Mentioned

BETA
reverse transcription PCR
RNA-seq
density gradient centrifugation
electron microscopy
PCR
affinity purification

Software Mentioned

WU
Affymetrix
BLAST
Integrated Genome Browser ( IGB )
Integrated Genome Browser (
SIROCCO
TargetRNA

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