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
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Cling-E. coliLet’s get together: Quorum-sensing
Stephanie Lo, Perry Tsai
July 9, 2007
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Random peptide display and screening
+Transformation
Expression
MACS(target with
magnetic bead)
FACS(target with fluorescent tag)
LppOmpA
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Target (eg Nickel)
luxIluxI
OHHL
luxRluxR
Bringing bacteria together
yo‘sup
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Test constructs
luxR
OHHL
BBa_I13263
BBa_T9002
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Growth data for logphase determination
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Fluorescence response to OHHL induction
GFP
YFP
100nM OHHL
10nM OHHLNon-induced No luxR
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BioBricks, assemble!
LuxI generator
luxI
LuxR generator / LuxI receiver / Reporter
luxR
LuxI/LuxR generator and receiver
luxI/luxR
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Looking into new BioBricks
LuxI amplifier
LuxI amplifier and reporter
LuxR generators with different promoters
LuxI receiver with different reporter
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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
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Synthesized LppOmpA (sLO)
Original LppOmpA
ACTAGAThrArgMixed
BioBrick-assembled LppOmpA
ACTAGAThrArgMixed
Synthesized LppOmpA
ACTAGAThrArgMixed
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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