RAPID BACTERIAL VIRULENCE ANNOTATION FOR THE POST GENOMIC ER A

Enormous amounts of money have been and are continuing to be spent on the genome sequencing of many different pathogenic bacteria. Despite this most of this sequence remains with little or no biological annotation. In order to achieve this biological annotation it would require extensive animal experimentation, which is both expensive and ethically problematic. Indeed it is widely recognized that there is a need to reduce the amount of animal experimentation currently conducted. With this in mind we have developed a technology for whole-genome mapping of bacterial virulence factors using invertebrate hosts that negates the need for mammalian testing. Simply we challenge three different invertebrate animals (insect, neamtode and amoeba) with cosmid libraries from a fully sequenced pathogen. End sequencing of cosmids that adversly effect any of these hosts are aligned with the genome sequence thus identifying pathogenic genes as bounded by clusters of cosmids. This fundamentally new approach allows us to rapidly and inexpensively bridge the enormous knowledge gap that has arisen through intensive genome sequencing. The current grant does not propose to follow-up on each of the virulence factors identified in detail but to provide access to the results as a community wide resource for workers studying each of the three bacteria. Details of the screens will be posted on the web with open access and other researchers will be able to choose cosmids for further research. Alternatively, we will also enter into collaborations with such workers to investigate genes in specific cosmids using the mutants generated.