Screening for genes affecting defense and endurance

Retest no.No. of lines testedNo. of positive linesNo. of negative lines% positive lines
Initial test12311018.2
  • Over 1200 PBac insertion mutant lines were tested for their ability to survive a L. monocytogenes infection and the MTD for each line was determined. The mutant lines that exhibited the most extreme phenotypes with MTDs of day 1 and day 7 (∼100 mutants) were considered positive hits and tested further to eliminate any false positives. The candidate mutants were tested in three independent experiments, the death curves for each were compared to the wild-type control, and log-rank statistical analysis was done. Lines that exhibited a death rate with a P-value <0.05 for all three retests were considered positive mutants. After three rounds of retesting >80% of these mutants were considered false positives and a total of 18 mutants, all with increased sensitivity to infection, were identified as positive mutants that affect host susceptibility to L. monocytogenes infection.