ABSTRACT: Salmonella typhimurium 14028s Transposon library recovered after three consecutive rounds of growth to late log phase in M9 minimal medium (arabinose 0.4%) at 37°C with aeration, compared to an expansion of the initial library selected on Luria agar plates + kanamycin (50ug/ml), O/N at 37°C Keywords: Transposon tag analysis 2 biological replicates analysed after arbitrarily primed sampling (Klenow, DOPR degenerate primer) + specific PCR amplification (Taq DNA Pol) + T7 in vitro transcription (AmpliScribe T7) + RT labeling (Cy3) + hybridization, compared to identically processed standard library (Cy5)
Project description:Comparisons of overexpression of PreA (in wt or preAB mutant Salmonella) to Salmonella lacking preA or preAB in late log phase growth in LB preA was externally supplied to wt or preAB mutant Salmonella typhimurium on a pBAD18 plasmid, which was then induced by 10mM arabinose and grown to late log phase (OD0.74). Transcription profiles were compared with those obtained from preA or preAB mutant Salmonella harboring pBAD18 only as vector control.