Cling-E. coli Let’s get together: Quorum-sensing

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

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

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

Test constructs OHHL luxR BBa_I13263 BBa_T9002

Growth data for log phase determination

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

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

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

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

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

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