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Diagnosing bacterial Diagnosing bacterial Diagnosing bacterial Diagnosing bacterial infections with viruses infections with viruses infections with viruses infections with viruses (Modular Construction of Bacteriophage for Diagnostic


  1. Diagnosing bacterial Diagnosing bacterial Diagnosing bacterial Diagnosing bacterial infections with viruses infections with viruses infections with viruses infections with viruses (Modular Construction of Bacteriophage for Diagnostic Systems) (Modular Construction of Bacteriophage for Diagnostic Systems) (Modular Construction of Bacteriophage for Diagnostic Systems) (Modular Construction of Bacteriophage for Diagnostic Systems) BIRKBECK BIRKBECK BIRKBECK BIRKBECK iGEM iGEM iGEM iGEM 2015 2015 2015 2015 A. Mirzarafie-Ahi, E. Parris, S. Craig, B. Steijl, L. Prout

  2. We are We are We are We are Birkbeck Birkbeck Birkbeck, University of London Birkbeck , University of London , University of London , University of London “London’s evening university” About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  3. Meet the Meet the Meet the Meet the Owligos Owligos Owligos Owligos About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  4. Infectious diseases in the developing world Infectious diseases in the developing world Infectious diseases in the developing world Infectious diseases in the developing world Infectious disease Treatment Detection Speed Cost About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  5. Case study: TB and antibiotic resistance Case study: TB and antibiotic resistance Case study: TB and antibiotic resistance Case study: TB and antibiotic resistance Source: World Health Organisation (2012) Images: World Health Organisation About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  6. Dr. Sanjib Bhakta Director of Mycobacteria Research Laboratory at Birkbeck/UCL What do you consider to be the main challenges in the eradication of TB and drug resistant strains? About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  7. About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  8. Dr. Muhammad Jawad Khan The Brooke Hospital Pakistan Are there any problems you frequently encounter with diagnosis? About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  9. About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  10. Fighting infectious disease with viruses Fighting infectious disease with viruses Fighting infectious disease with viruses Fighting infectious disease with viruses “The enemy of my enemy is my friend” Tail Short tail E. coli fibres Baseplate Lambda phage About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  11. Fighting infectious disease with viruses Fighting infectious disease with viruses Fighting infectious disease with viruses Fighting infectious disease with viruses Fighting infectious disease with viruses Fighting infectious disease with viruses Fighting infectious disease with viruses Fighting infectious disease with viruses 4.5 h TB Jacobs et al (2012) Journal of Clinical Microbiology , 50(4), 1362; doi: 10.1128/JCM.06192-11 About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  12. Fighting infectious disease with viruses Fighting infectious disease with viruses Fighting infectious disease with viruses Fighting infectious disease with viruses 1. Patient provides 2. Sample mixed with 3. Signal in presence sample phage solution of pathogen GFP About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  13. Fighting infectious disease with viruses Fighting infectious disease with viruses Fighting infectious disease with viruses Fighting infectious disease with viruses 1. Patient provides 2. Sample mixed with 3. Signal in presence sample phage solution of pathogen GFP etc. About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  14. Plan of Attack Plan of Attack – Plan of Attack Plan of Attack – – – Developing Modularity Developing Modularity Developing Modularity Developing Modularity using Bacteriophage lambda (arch enemy of E.coli ) as chassis phage tail fibre genome assembly protein (tfa) chaperone – facilitates phage tail fibre protein folding protein shell tail fibre tip attachment protein (stf) facilitates initial attachment protein J of virion to E.coli outer binds to E.coli membrane protein C maltoporin (LamB) (OmpC) About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  15. Directed Evolution of Short Tail Directed Evolution of Short Tail Fibres Directed Evolution of Short Tail Directed Evolution of Short Tail Fibres Fibres Fibres lambda phage short tail fibres E. coli mutant E. coli receptor About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  16. Directed Evolution of Short Tail Directed Evolution of Directed Evolution of Directed Evolution of Short Tail Short Tail Fibres Short Tail Fibres Fibres Fibres lambda phage short tail fibres E. coli mutant E. coli receptor About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  17. Directed Evolution of Directed Evolution of Short Tail Directed Evolution of Directed Evolution of Short Tail Short Tail Fibres Short Tail Fibres Fibres Fibres engineered lambda phage short tail fibres E. coli receptor E. coli mutant About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  18. Changing bacteriophage specificity Changing bacteriophage specificity Changing bacteriophage specificity Changing bacteriophage specificity Circuit Development About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  19. Changing bacteriophage specificity Changing bacteriophage specificity Changing bacteriophage specificity Changing bacteriophage specificity Circuit Development About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  20. Changing bacteriophage specificity Changing bacteriophage specificity Changing bacteriophage specificity Changing bacteriophage specificity Circuit Development Figure 1. Analysis of cI/Cro circuit in DH10β E. coli alongside control samples showing successful expression of the cI repressor under a constitutive promoter and no expression in T7 Express E. coli cells. 1, 2, 3: T7 Express E. coli 4, 5, 6: T7 Express E. coli + cl-Crocircuit 7, 8, 9: DH10β E. coli + cl-Cro circuit 10, 11: DH10β E. coli + with IPTG (0.4 µg/ml [final]) ‡ with IPTG (2.0 µg/ml [final]) About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  21. Changing bacteriophage specificity Changing bacteriophage specificity Changing bacteriophage specificity Changing bacteriophage specificity Circuit Development About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  22. Changing bacteriophage specificity Changing bacteriophage specificity Changing bacteriophage specificity Changing bacteriophage specificity Circuit Development Figure 2. Analysis of TetR-regulated tfa (tail fibre assembly) circuit in DH10β E. coli alongside control samples showing successful expression of the tfa protein under TetR regulation. 1, 2, 3: tfa composite part (control) 4, 5, 6: TetRcomposite part (control) 7, 8, 9: TetR-regulated tfa circuit in pSΒ1A3 10: DH10β (control) 11, 12, 13: TetR-regulated tfa circuit in pSΒ1K3 *with anhydrous tetracycline (aTc) (0.4 µg/ml [final]) † with aTc (2.0 µg/ml [final]) About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  23. Future Work: Future Work: Future Work: Future Work: Changing bacteriophage specificity Changing bacteriophage specificity Changing bacteriophage specificity Changing bacteriophage specificity Directed Evolution Strategy About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  24. New Parts submitted to the Registry New Parts submitted to the Registry New Parts submitted to the Registry New Parts submitted to the Registry basic “modular” parts basic “modular” parts basic “modular” parts basic “modular” parts Name Construct Description Status Part Goal Basic part ORF-314 BBa_K1846000 Complete stf Basic part BBa_K1846004 Complete gene tfa Basic part in production gene J gene Basic part in production About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

  25. New Parts submitted to the Registry New Parts submitted to the Registry New Parts submitted to the Registry New Parts submitted to the Registry composite parts and circuits composite parts and circuits composite parts and circuits composite parts and circuits Name Construct Description Status Part Goal Composite tfa tail fibre assembly part BBa_K1846001 construct construct Complete P(Cat)- Composite construct for the TetR part BBa_K1846003 production of TetR Complete construct Composite TetR-tfa circuit to control tail fibre part BBa_K1846007 circuit assembly protein Complete circuit to control lytic and Composite cI-cro lysogenic cycles of part BBa_K1846005 circuit lambda phage Complete About us - Background - Plan of attack - BioBricks - Conclusions - Acknowledgements

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