Making delayed-LuxR mutants

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Experiments

  1. luxR gene was mutated by error-prone PCR using Mn2+(Joyce, 1996) and cloned it into a pUC vector.
  2. This library was transformed into XL10-Gold, and the plasmid was mini-prepped to get an plasmid library (pUC-[luxR]).
  3. The plasmid library was transformed into JW1226 harboring pAC-plux-gfp, which will glow green when an active LuxR was present.
  4. We screened for the fast/dull mutant of LuxR, and collected the clone's plasmid.
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