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Analysis of gene expression profiles allowing zoonotic Brucella species to adapt to their intracellular niche and to environmental stress
Project
Project code: BfR-BIOS-08-1329-452
Contract period: 01.12.2010
- 30.09.2012
Purpose of research: Applied research
Brucellosis is a globally re-emerging zoonosis. The infection is caused by a variety of different Brucella species which are genetically closely related but phenotypically divergent. There are crucial differences in environmental persistence, pathogenicity and host specificity of the pathogens. Gene regulation and metabolic adaptation might be responsible for the apparent contradiction between genotype and phenotypic traits.
In the research project, a comparative transcriptome analysis of the two homologous species B. suis and B. microti (99.84% genome sequence identity) shall be performed under nutrient- and pH-stress in vitro, mimicking starvation and acidic conditions in the host and the environment. To this end, bacteria will be cultivated in defined minimal media at pH 7 and pH 4.5. B. microti has been recently described as a new member of the genus Brucella and the first species that was isolated from habitats outside mammalian hosts. The isolation of B. microti from soil samples represents an unpredictable risk for men and animals, as this reservoir may be linked to so far unknown modes of transmission. B. microti proved to be more resistant to acidic environments than the classical Brucella species so that the bacteria may survive cheese ripening and persist in food products despite conservation and long-term storage. In addition, this new species was highly virulent in the murine model of infection.
In the planned study, the gene expression profiles will be analyzed using high-throughput total RNA sequencing („RNA Seq'; Illumina/Solexa). In contrast to DNA-microarray hybridization techniques, the transcripts are analyzed without a priori knowledge of their sequences, a broad spectrum of different expression levels may be detected and all transcribed regions of the genome can be mapped. Data analysis will be followed by biological validation based on qPCR and other genetic methods.
Last but not least, Brucella will act as a model pathogen for bacterial zoonotic diseases with the distinctive feature that individual species are restricted to specific animal hosts, despite highly conserved genomes. The identification of specific regulatory processes and networks therefore appears promising and of great impact on the understanding of host specificity and adaptation to environmental stress conditions. The expected results of the study will complement our knowledge deriving from metabolic activity testing of Brucella spp. which is currently conducted at the Federal Institute for Risk Assessment.
Section overview
Subjects
- Animal health
- Biotechnology
