1. Lambda phage killing: infection and “parts” N Kuldell for 20.020 Spring 2009

2. Heidelburg iGEM 2008: ecolicense to kill Part 2: DNA transfer, genetically programmed self-assembly and “parts” Background information

3. DNA transfer natural context: 3 mechanisms http://www.scq.ubc.ca/wp-content/uploads/2006/08/HorizontalTransfer.gif

4. “Transduction” by bacteriophage www.phagebiotech.com/ Protein coat encapsulates nucleic acids Bacteriophage: viruses that infect bacteria INFECT REPLICATE ASSEMBLE RELEASE

5. “Temperate” bacteriophage lambda http://escience.ws/b572/L17/L17.htm INFECT REPLICATE ASSEMBLE RELEASE lytic

6. http://escience.ws/b572/L17/L17.htm INFECT REPLICATE ASSEMBLE RELEASE lytic lysogenic “Temperate” bacteriophage lambda

7. http://escience.ws/b572/L17/L17.htm INFECT REPLICATE ASSEMBLE RELEASE “Temperate” bacteriophage lambda Some great ??s What guides lytic/lysogenic decision? What keeps lysogen stable? What triggers lysogen to lytic cycle?

8. http://escience.ws/b572/L17/L17.htm INFECT REPLICATE ASSEMBLE RELEASE “Temperate” bacteriophage lambda Some great ??s the Heidelberg team asked: Can we infect prey from lysogen (=predator)? Can we keep lysogen from lysing itself? Can we monitor lysis and lysogeny?

9. http://escience.ws/b572/L17/L17.htm INFECT REPLICATE ASSEMBLE RELEASE “Temperate” bacteriophage lambda Some great ??s the Heidelberg team asked: Can we infect prey from lysogen (=predator)? Can we keep lysogen from lysing itself? Can we monitor lysis and lysogeny?

10. Genetically programmed infection iGEM context: phage infection via conjugation

11. Genetically programmed infection iGEM context: phage infection via conjugation http://escience.ws/b 572/L18/L18.htm io.uwinnipeg. ca

12. Genetically programmed infection iGEM context: phage infection via conjugation What “parts” are needed Pilus…. “F’” Origin for transfer… “oriT” Selectable marker… “CamR” Model predicts: 10 killer cells kill 10^9 prey cells “in silico” F’ oriT CamR “Part” is a genetically-encoded, human defined function

13. Genetically programmed infection iGEM context: phage infection via conjugation What “parts” are needed Pilus…. “F’” Origin for transfer… “oriT” Selectable marker… “CamR” F’ oriT CamR “Part” is a genetically-encoded, human defined function I714030

14. http://escience.ws/b572/L17/L17.htm INFECT REPLICATE ASSEMBLE RELEASE “Temperate” bacteriophage lambda Some great ??s the Heidelberg team asked: Can we infect prey from lysogen (=predator)? Can we keep lysogen from lysing itself? Can we monitor lysis and lysogeny?

15. http://escience.ws/b572/L17/L17.htm INFECT REPLICATE ASSEMBLE RELEASE “Temperate” bacteriophage lambda Some great ??s the Heidelberg team asked: Can we infect prey from lysogen (=predator)? Can we keep lysogen from lysing itself? Can we monitor lysis and lysogeny?

16. Genetically programmed bi-stable switch natural context: epigenetic regulation Genes Dev. (2004)18(17): 2086–2094. doi: 10.1101/gad.1226004. lysogeny lysis

17. Genetically programmed bi-stable switch iGEM context: flip and hold in one state Genes Dev. (2004)18(17): 2086–2094. doi: 10.1101/gad.1226004. lysogeny promoterRBScI ORF transcriptional terminator

18. http://escience.ws/b572/L17/L17.htm INFECT REPLICATE ASSEMBLE RELEASE “Temperate” bacteriophage lambda Some great ??s the Heidelberg team asked: Can we infect prey from lysogen (=predator)? Can we keep lysogen from lysing itself? Can we monitor lysis and lysogeny?

19. http://escience.ws/b572/L17/L17.htm INFECT REPLICATE ASSEMBLE RELEASE “Temperate” bacteriophage lambda Some great ??s the Heidelberg team asked: Can we infect prey from lysogen (=predator)? Can we keep lysogen from lysing itself? Can we monitor lysis and lysogeny?

20. Fluorescence vs bioluminescence AI-2 AI-1

21. Fluorescence vs bioluminescence AI-2 AI-1 http://www.lifesci.ucsb.edu/~biolum/chem/

22. Fluorescence vs bioluminescence AI-2 AI-1 http://www.lifesci.ucsb.edu/~biolum/chem/

23. One last thing about fluorescence

24. Genetically programmed infection iGEM context: phage infection via conjugation What “parts” are needed Pilus…. “F’” Origin for transfer… “oriT” Selectable marker… “CamR” Stable lysogen… “cI” Moniter… “GFP” F’ oriT CamR

25. Predator cell stable lysogen that infects by conjugation + CamR + GFP to follow process