Abstract

Integrity of prokaryotic mRNA isolated from complex samples for in vivo bacterial transcriptome analysis

Author(s): A.B. Ferreira-Machado, M.C.R. Freitas, G.R.Q. Saji, A.B. Rezende, P.E. Almeida, D.E. Cesar, J.A. Resende1, M.F. Nic�³las2, V.L. Silva and C.G. Diniz

Even though several in vitro studies have focused on bacterial biology, the extent of such knowledge is not complete when considering an actual infection. As culture-independent microbiology methods such as high-throughput sequencing became available, important aspects of host-bacterium interactions will be elucidated. Based on microbiological relevance, we considered Bacteroides fragilis in a murine experimental infection as a model system to evaluate the in vivo bacterial transcriptome in host exudates. A disproportionate number of reads belonging to the host genome were retrieved in the first round of pyrosequencing, even after depletion of ribosomal RNA;

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