Quorum Sensing: Week 6 Results

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Presentation transcript:

Quorum Sensing: Week 6 Results Two Cell vs. One Cell Systems

Switched Fluorophores Original Constructs Senders lacI regulated RFP Sender (S08I07) tetR regulated RFP Sender (S23I07) Receiver GFP Receiver (T02)

Switched Fluorophores Flipped Constructs Senders lacI regulated GFP Sender (S08E40) tetR regulated GFP Sender (S23E40) Receiver RFP Receiver (F20I07)

Switched Fluorophores

Switched Fluorophores

Switched Fluorophores Protein Folding/Maturation issue? GFP (E0040) gfpmut3b “The expression of GFP is observable minutes after induction”1 RFP (E1010) mRFP1 - “**highly** engineered mutant of red fluorescent protein from Discosoma striata (coral)” “Based on our data at 37°C, 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 T1”2 1 “FACS-optimized mutants of the green fluorescent protein (GFP).”Gene [0378-1119] Cormack yr:1996 vol:173 iss:1 Spec No pg:33 -8 2 “A monomeric red fluorescent protein.” Proc Natl Acad Sci U S A. 2002 June 11; 99(12): 7877–7882. doi: 10.1073/pnas.082243699.

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.

FACS - Results 1:10 Dilution Final OD = 0.02 Mean GFP = 370