cling- e. coli
DESCRIPTION
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. - PowerPoint PPT PresentationTRANSCRIPT
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