抗生素耐药Engineering a virus to select against antibiotic resistancePPT.pptVIP

This is our plan of attack. First we outlined our circuit and used computational modeling to refine our design. Then we constructed multiple versions of the circuit and characterized them in plasmids. We plan on using the data from this characterization to further refine our modeling and design until the circuit is fully functional. Now Thomas will explain our circuit design and construction. Time (min) Excitation = 514 nm, Emission = 527 nm In contrast, EYFP is produced under similar conditions in TcS E. coli TcS+circuit TcS-circuit [aTc] = 1 μM KD = 0.01nM Biochemistry. Vol. 35:23 p.7439-7446 0 1 10 5 2 10 5 3 10 5 4 10 5 5 10 5 6 10 5 7 10 5 0 20 40 60 80 100 Fluorescence/A600 Effects of increasing Tc on cell growth [Tc] ≤ 2 μM can be used to drive our circuit with large adverse effects on cell growth However, the effects of Tc on EYFP (and CI) production have not yet been characterized. TcR-circuit TcR+circuit TcS-circuit TcS+circuit T = 240min Conclusions Computational Modeling ??created effective model and selected circuit for characterization Circuit Design Construction ??built a variety of selection circuits Circuit Characterization ??demonstrated circuit’s ability to differentiate between cell types Future Roadmap Computational Modeling ??model circuit in phage Circuit Design Construction ??construct phage optimized circuit Circuit Characterization ??test circuit function within phage COMPUTATION AND APPLIED MATH Steve Cox Jay Raol CHEMICAL AND BIOMOLECULAR ENGINEERING Ken Cox Alec Walker Baylor College of Medicine Bibhash Mukhopadhyay BIOCHEMISTRY AND CELL BIOLOGY Beth Beason George Bennett Tina Chen David Kim Joff Silberg Taylor Stevenson Arielle Layman BIOENGINEERING Christie Peebles Ka-Yiu San Thomas-Segall Shapiro Linking our cell detection to phage lysis 0 1 10 5 2 10 5 3 10 5 4 10 5 5 10 5 6 10 5 7 10 5 0 20 40 60 80 Fluorescence/A600 Time (min) Time (min) 0 20 40 60 80 Create TcR and TcS temperature se