cling- e. coli

Cling-E. coliLet’s get together: Quorum-sensing

Stephanie Lo, Perry Tsai

July 9, 2007

Random peptide display and screening

+Transformation

Expression

MACS(target with

magnetic bead)

FACS(target with fluorescent tag)

LppOmpA

Target (eg Nickel)

luxIluxI

OHHL

luxRluxR

Bringing bacteria together

yo‘sup

Test constructs

luxR

OHHL

BBa_I13263

BBa_T9002

Growth data for logphase determination

Fluorescence response to OHHL induction

GFP

YFP

100nM OHHL

10nM OHHLNon-induced No luxR

BioBricks, assemble!

LuxI generator

luxI

LuxR generator / LuxI receiver / Reporter

luxR

LuxI/LuxR generator and receiver

luxI/luxR

Looking into new BioBricks

LuxI amplifier

LuxI amplifier and reporter

LuxR generators with different promoters

LuxI receiver with different reporter

In other LppOmpA news… Lpp-OmpA PDZ Western blot

(+) LO-PDZ IPTG(-)

LO-PDZ IPTG(+)

0 1 2 1+2 0 1 2 1+2

GFP-Mek

PDZ domain

Synthesized LppOmpA (sLO)

Original LppOmpA

ACTAGAThrArgMixed

BioBrick-assembled LppOmpA

ACTAGAThrArgMixed

Synthesized LppOmpA

ACTAGAThrArgMixed

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