Vibrio cholerae Transcriptome
Vibrio cholerae is an intestinal pathogen that causes cholera, a severe and sometime lethal diarrheal disease. V. cholerae is a Gram-negative facultative pathogen that can inhabit two distinct niches; this highly motile bacterium lives in the natural aquatic reservoir and can also survive and multiply in the human intestine, causing devastating illness. A primary mechanism by which the organism adapts to these different environments is through changes in global patterns of transcription. These changes alter the organism?s metabolic and protein repertoire. Thus, information about global changes in the V. cholerae ?transcriptome? in different in vitro as well as in vivo conditions will be invaluable to enable analyses of these adaptive processes and provide potential insight for therapeutics.
The current information about the V. cholerae transcriptome comes from recombination based in vivo technology and microarray analyses. The first technique is extremely laborious and is very low throughput and does not allow for comprehensive global analyses of the transcriptome. Microarrays and other hybridization-based techniques have several important. In nearly all cases, microarrays do not contain complete representations of both strands of the entire genome. For example, small noncoding RNA species, which are often encoded in ?intergenic? regions, are usually not present in microarrays. Furthermore, most microarrays do not contain DNA that corresponds to ?antisense? transcripts. However, recent studies suggest that antisense transcription may be common. In addition, microarrays do not allow for identification of primary versus processed transcripts. Finally, cross-hybridizations can limit the specificity of microarray studies.
The development of next-generation sequencing technology and RNA sequencing is enabling a new level of depth and accuracy in high-throughput transcriptome analyses, without the above mentioned drawbacks. We propose to utilize Illumina next-generation RNA sequencing technology to define the complete V. cholerae transcriptome under a variety of conditions to further our understanding of how V. cholerae adapts to the varied niches it can inhabit.
Samples are in the queue for sequencing, check back for data.
This sequencing project was supported by the National Institute of Allergy and Infectious Disease, National Institutes of Health funded Genome Sequencing Center for Infectious Diseases at the Broad Institute.