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Quorum Sensing: Week 6 Results
Two Cell vs. One Cell Systems
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Switched Fluorophores
Original Constructs Senders lacI regulated RFP Sender (S08I07) tetR regulated RFP Sender (S23I07) Receiver GFP Receiver (T02)
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Switched Fluorophores
Flipped Constructs Senders lacI regulated GFP Sender (S08E40) tetR regulated GFP Sender (S23E40) Receiver RFP Receiver (F20I07)
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Switched Fluorophores
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Switched Fluorophores
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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 [ ] 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 June 11; 99(12): 7877–7882. doi: /pnas
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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):
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FACS - Results 1:10 Dilution Final OD = 0.02 Mean GFP = 370
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