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The exploration of Brucella transcriptome: From the ORFeome to RNAseq

AuthorsGarcía-Lobo, Juan M. CSIC ORCID; Rodríguez, María Cruz CSIC; Seoane, Asunción CSIC; Sangari, Félix J. CSIC ORCID; López-Goñi, Ignacio
Issue Date2012
PublisherCaister Academic Press
CitationBrucella: Molecular Microbiology and Genomics (Cap.5): 89-102 (2012)
AbstractIn this chapter we will analyze the results available on the characterization of the Brucella transcriptome. After a summary of earlier work on transcription, two technical approaches will be mainly described, on one side the use microarrays, specially that derived from the Brucella ORFeome that allows hybridization with mRNA derived cDNA to determine the relative abundance of transcripts from each B. melitensis ORF. On the other, RNAseq, consisting in the massive sequencing of cDNA libraries derived from mRNA obtained from B. abortus grown in culture medium. Sequencing with the Illumina Genome-Analyzer II platform, produced 3 millions of 35 nt long reads that annealed with single copy coding regions of the genome. This allowed a good coverage for every CDS and produced a new dataset on the transcription of Brucella. We obtained a good correlation for the set of highly expressed genes from the microarrays and confirmed the observations obtained on the asymmetry between chromosome transcription. Preliminary conclusions on intracellular transcription have been drawn from RT-PCR on selected candidate genes and from microarray datasets obtained from virulence related conditions.The RNAseq derived data allowed more versatile data mining giving some new details on transcription from pseudogenes or intergenic regions.
DescriptionEl pdf es la versión post-print.
Identifiersisbn: 978-1-904455-93-6
Appears in Collections:(IBBTEC) Libros y partes de libros

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