abstract book - Clostridia

GENETIC ANALYSIS OF CLOSTRIDIUM PERFRINGENSJ.I. Rood, D. Lyras, J.K. Cheung, T.L. Bannam & M. M. AwadAustralian Bacterial Pathogenesis Program and ARC Centre of Excellencein Structural and Functional Microbial Genomics, Department ofMicrobiology, Monash University, Clayton, Vic 3800, Australia.Clostridium perfringens is undoubtedly the pathogenic clostridium mostamenable to genetic manipulation. In addition, unlike any other clostridium,there are three complete, annotated genome sequences available. For manyyears it has been possible to electroporate some virulent isolates at afrequency that is high enough to allow for the construction of chromosomalmutants by double crossover events. Unfortunately, many other strains arestill not transformable. There is a well established series of shuttle vectorsavailable for complementation studies, including shuttle plasmids that can beused to introduce genes into C. perfringens by RP4-dependent conjugativetransfer from Escherichia coli. Finally, group II intron-based technology canalso be used to construct defined chromosomal mutants in C. perfringens.We have used various combinations of these genetic tools to analyse thefunctional role of toxins in disease, to determine how these toxin genes areregulated and to carry out genetic analysis of the conjugation process in C.perfringens.