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

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Presentation on theme: "Cling-E. coli Let’s get together: Quorum-sensing"— Presentation transcript:

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 LppOmpA MACS (target with magnetic bead) FACS (target with fluorescent tag)

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

4 Test constructs OHHL luxR BBa_I13263 BBa_T9002

5 Growth data for log phase determination

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

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

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
GFP-Mek PDZ domain (+) LO-PDZ IPTG(-) IPTG(+)

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