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Dual reporters in bacterial pathogens Philip J Hill*; Saara NA Qazi; Catherine ED Rees University of Nottingham, School of Biosciences, Sutton Bonington Campus, Sutton Bonington, Leics, LE12 5RD, UK      *(phil.hill@nottingham.ac.uk) Bacterial Lux and fluorescent proteins such as GFP have their own particular advantages and disadvantages as reporter/marker genes. The major disadvantages of Lux are the requirement for live cells and the poor spatial resolution when using low light imaging microscopy. These disadvantages represent two of the major advantages of the fluorescent proteins, which, unlike lux, suffers from poor sensitivity and high-background fluorescence problems. Clearly the utilisation of both of these reporters represents an opportunity to capitalise on the complementary strengths of each. To this end we have constructed novel dual reporter operons for high level expression in Gram positive and Gram negative bacteria, incorporating a modified lux operon in conjunction with dsRED or GFP variants of differing half-lives. These have been used for tracking of bacteria and evaluated as reporters of gene expression in Gram positive pathogens. This new system allows bacterial location and gene expression to be assessed both in “real-time” in vivo, and also “retrospectively” on fixed samples. [Talk: hill.philip.86081]

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