Engineered E. coli Strain for Versatile Genetic Manipulation

All DNA cloning was performed with Mach1 cells (Invitrogen) or NEB Turbo cells (New England Biolabs). All discrete infection assays, plaque assays and PACE experiments were performed with E. coli S1030. This strain was derived from E. coli S109^{1 (link)} and was modified using the Lambda Red method^{23 (link)} as follows: 1) scarless mutation to rpoZ to introduce a frameshift mutation to enable n-hybrid schemes;^{24 (link)} 2) integration of lacI and tetR overexpression cassettes onto the F plasmid to enable small-molecule regulated transcription of various genes; 3) integration of luxCDE onto the F plasmid for the production of decanal to facilitate luciferase monitoring experiments;^{25 (link)} 4) deletion of flu/pgaC/csgABCDEFG to dramatically reduce biofilm formation in chemostat PACE experiments;^{26 (link)-28 (link)} and 5) mutation of the chromosomal, low-affinity high-capacity AraE promoter to a constitutive promoter for titratable arabinose induction of the P_BAD promoter on the mutagenesis plasmid.^{29 (link)} The complete genotype of the resulting strain is F'proA+B+ Δ(lacIZY) zzf∷Tn10(Tet^R) lacI^Q1 P_N25-tetR luxCDE/endA1 recA1 galE15 galK16 nupG rpsL(Str^R) ΔlacIZYA araD139 Δ(ara,leu)7697 mcrA Δ(mrr-hsdRMS-mcrBC) proBA∷pir116 araE201 ΔrpoZ Δflu ΔcsgABCDEFG ΔpgaC λ⁻.