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Lambda phage killing: infection and “parts”

Lambda phage killing: infection and “parts”. N Kuldell for 20.020 Spring 2009. Heidelburg iGEM 2008: ecolicense to kill. Part 2: DNA transfer, genetically programmed self-assembly and “parts”. Background information. DNA transfer natural context: 3 mechanisms.

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Lambda phage killing: infection and “parts”

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

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

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

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

  8. INFECT REPLICATE RELEASE ASSEMBLE “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? http://escience.ws/b572/L17/L17.htm

  9. INFECT REPLICATE RELEASE ASSEMBLE “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? http://escience.ws/b572/L17/L17.htm

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

  11. Genetically programmed infection iGEM context: phage infection via conjugation io.uwinnipeg.ca http://escience.ws/b572/L18/L18.htm

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

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

  14. INFECT REPLICATE RELEASE ASSEMBLE “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? http://escience.ws/b572/L17/L17.htm

  15. INFECT REPLICATE RELEASE ASSEMBLE “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? http://escience.ws/b572/L17/L17.htm

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

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

  18. INFECT REPLICATE RELEASE ASSEMBLE “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? http://escience.ws/b572/L17/L17.htm

  19. INFECT REPLICATE RELEASE ASSEMBLE “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? http://escience.ws/b572/L17/L17.htm

  20. AI-2 AI-1 Fluorescence vs bioluminescence

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

  22. AI-2 AI-1 Fluorescence vs bioluminescence 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” CamR oriT F’

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

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