抗生素耐药Engineering a virus to select against antibiotic resistance课件.ppt

Ptet Controlled Production of TetR Creates Resistance Phenotype 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 sensitive lysogens and determine if circuit provides a relative fitness gain for TcS over TcR cells TcR+/-circuit TcS+/-circuit TcS+circuit tetR Ptet TcS I744204 B0011 Tetracycline Affects Growth Rates t = 30min 1744204 B0011 (empty) Tetracycline Affects on Growth Rates t = 240min TcR TcR+circuit TcS TcS+circuit I744204 B0011 Phage C