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Exploiting complete genome sequence data to develop genetic tools for analysis of the obligately anaerobic bacterium Bacteroides fragilis

Reference	BB/C505875/1
Principal Investigator / Supervisor	Dr Garry Blakely
Co-Investigators / Co-Supervisors
Institution	University of Edinburgh
Department	Inst of Cell Biology
Funding type	Research
Value (£)	120,260
Status	Completed
Type	Research Grant
Start date	01/02/2005
End date	31/01/2007
Duration	24 months

Abstract

The Gram negative strictly anaerobic Bacteroidetes represent one of the predominant bacterial groups within the resident microbiota of the human gastrointestinal tract. Bacteroides fragilis is an important member of the normal gut microbiome but is also responsible for causing clinical infections. Capsular polysaccharides on the surface of the bacterial cell are variable and presumably have a role in survival of the organism in the host. The genome sequence of B. fragilis has recently been completed and has demonstrated a remarkable plasticity. An unprecedented number of invertible promoter regions upstream of genes involved in the production of surface and secreted components, are responsible for gene regulation. In addition a number of long-range genome rearrangements, ranging from 5-100 kb, have also been identified. Genetical studies of B. fragilis have been hampered by the lack of appropriate tools. The genome sequence has now revealed that B. fragilis contains numerous restriction/modification systems. We will develop new methods, using DNA mimics, that will inhibit restriction enzymes in B. fragilis and overcome the barrier to introduction of unmodified DNA. We will also develop new tools, using site-specific recombination, that will enable generation of non-polar deletions to allow the study of gene function within operons. These tools will be used to determine if two conserved paralogous families of potential regulatory proteins are involved in the transcriptional control of polysaccharide biosynthetic operons. Initial analysis of long-range DNA inversions will concentrate on identifying the recombinase responsible for a specific inversion on the chromosome. A better understanding of this bacterium will help us to appreciate its role in the normal development of the gut microbiota and will enable the future development of novel antimicrobial agents.

Summary

unavailable

Committee	Closed Committee - Plant & Microbial Sciences (PMS)
Research Topics	X – not assigned to a current Research Topic
Research Priority	X – Research Priority information not available
Research Initiative	X - not in an Initiative
Funding Scheme	X – not Funded via a specific Funding Scheme

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