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Polyamine biosynthesis is not required for fitness in serum ex vivo.

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posted on 22.04.2019, 17:40 by Chelsie E. Armbruster, Valerie S. Forsyth, Alexandra O. Johnson, Sara N. Smith, Ashley N. White, Aimee L. Brauer, Brian S. Learman, Lili Zhao, Weisheng Wu, Mark T. Anderson, Michael A. Bachman, Harry L. T. Mobley

50% naive mouse serum was inoculated with 1x106 CFU wild-type P. mirabilis, an isogenic mutant, or a 1:1 mixture of wild-type P. mirabilis and isogenic mutant (co-culture). The following mutants that exhibited significant fitness defects in the liver and/or spleen during bacteremia were included: speA (A), speB (C), and potB (E). Cultures were incubated statically at 37°C with 5% CO2 and sampled at 0, 1, 3, 6, and 24 hours for enumeration of CFUs on LB agar with and without kanamycin. Error bars represent the mean ± standard deviation from three biological replicates. (B, D, F) A competitive index (CI) was calculated for the mutant at each time point from the ratio of mutant to wild-type colonization level at the time indicated divided by the ratio of mutant to wild-type at time zero (see Materials and Methods). Each data point represents the CI from an individual replicate. Solid lines represent the median. Dashed lines indicate a competitive index of 1, or a 1:1 ratio of mutant to wild-type. Error bars represent the mean ± standard deviation from two or three biological replicates.