Title: Quorum Sensing: Week 6 Results
1Quorum Sensing Week 6 Results
- Two Cell vs. One Cell Systems
2Switched Fluorophores
Original Constructs
Senders
lacI regulated RFP Sender (S08I07)
tetR regulated RFP Sender (S23I07)
Receiver
GFP Receiver (T02)
3Switched Fluorophores
Flipped Constructs
Senders
lacI regulated GFP Sender (S08E40)
tetR regulated GFP Sender (S23E40)
Receiver
RFP Receiver (F20I07)
4Switched Fluorophores
5Switched Fluorophores
6Switched Fluorophores
- Protein Folding/Maturation issue?
- GFP (E0040)
- gfpmut3b
- The expression of GFP is observable minutes
after induction1 - RFP (E1010)
- mRFP1 - highly engineered mutant of red
fluorescent protein from Discosoma striata
(coral) - Based on our data at 37C, we estimate the t0.5
for maturation of mRFP1 and T1 is less than 1 h,
consistent with the 0.7 h previously reported for
T12
1 FACS-optimized mutants of the green
fluorescent protein (GFP).Gene 0378-1119
Cormack yr1996 vol173 iss1 Spec No pg33 -8 2
A monomeric red fluorescent protein. Proc Natl
Acad Sci U S A. 2002 June 11 99(12) 78777882.
doi 10.1073/pnas.082243699.
7 FACS - Inspiration
Anderson, J. C., Clarke, E. J., Arkin, A. P.,
Voigt, C. A. (2006) Environmentally Controlled
Invasion of Cancer Cells by Engineered Bacteria,
Journal of Molecular Biology, 355 (4) 619-627.
8 FACS - Results
110 Dilution Final OD 0.02 Mean GFP 370
1100 Dilution Final OD 0.141 Mean GFP 315
11000 Dilution Final OD 0.555 Mean GFP 426