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Cling- E. coli

Cling- E. coli. Let’s get together: Quorum-sensing Stephanie Lo, Perry Tsai July 9, 2007. Random peptide display and screening. Transformation Expression. +. FACS (target with fluorescent tag). MACS (target with magnetic bead). LppOmpA. Target (eg Nickel). luxR. luxI. luxI. luxR.

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Cling- E. coli

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  1. Cling-E. coli Let’s get together: Quorum-sensing Stephanie Lo, Perry Tsai July 9, 2007

  2. Random peptide display and screening Transformation Expression + FACS (target with fluorescent tag) MACS (target with magnetic bead) LppOmpA

  3. Target (eg Nickel) luxR luxI luxI luxR yo ‘sup OHHL Bringing bacteria together

  4. OHHL luxR Test constructs BBa_I13263 BBa_T9002

  5. Growth data for logphase determination

  6. Fluorescence response to OHHL induction GFP YFP 100nM OHHL 10nM OHHL Non-induced No luxR

  7. BioBricks, assemble! luxR luxI LuxI generator LuxR generator / LuxI receiver / Reporter luxI/luxR LuxI/LuxR generator and receiver

  8. Looking into new BioBricks LuxI amplifier LuxR generators with different promoters LuxI amplifier and reporter LuxI receiver with different reporter

  9. In other LppOmpA news… Lpp-OmpA PDZ Western blot PDZ domain (+) LO-PDZ IPTG(-) LO-PDZ IPTG(+) GFP-Mek 0 1 2 1+2 0 1 2 1+2

  10. Synthesized LppOmpA (sLO) Original LppOmpA BioBrick-assembled LppOmpA ACTAGA ThrArg Mixed ACTAGA ThrArg Mixed Synthesized LppOmpA ACTAGA ThrArg Mixed

  11. This week’s agenda • FACS appointment on July 11 • Checking out ligations and new BioBricks • More fluorescence assays, possibly with new constructs • Construction of GFPmek for PDZ binding • Preparing sLppOmpA for future assembly

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