Lambda phage killing infection and parts N Kuldell

Lambda phage killing: infection and “parts” N Kuldell for 20. 020 Spring 2009

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

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

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

“Temperate” bacteriophage lambda INFECT lytic REPLICATE RELEASE ASSEMBLE http: //escience. ws/b 572/L 17. htm

“Temperate” bacteriophage lambda INFECT lytic REPLICATE RELEASE lysogenic ASSEMBLE http: //escience. ws/b 572/L 17. htm

“Temperate” bacteriophage lambda Some great ? ? s What guides lytic/lysogenic decision? What keeps lysogen stable? What triggers lysogen to lytic cycle? INFECT REPLICATE RELEASE ASSEMBLE http: //escience. ws/b 572/L 17. htm

“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? INFECT REPLICATE RELEASE ASSEMBLE http: //escience. ws/b 572/L 17. htm

“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? INFECT REPLICATE RELEASE ASSEMBLE http: //escience. ws/b 572/L 17. htm

Genetically programmed infection i. GEM context: phage infection via conjugation

Genetically programmed infection i. GEM context: phage infection via conjugation io. uwinnipeg. ca http: //escience. ws/b 572/L 18. htm

Genetically programmed infection i. GEM context: phage infection via conjugation What “parts” are needed Pilus…. “F’” Origin for transfer… “ori. T” Selectable marker… “Cam. R” “Part” is a genetically-encoded, human defined function Model predicts: 10 killer cells kill 10^9 prey cells “in silico” Cam. R ori. T F’

Genetically programmed infection i. GEM context: phage infection via conjugation What “parts” are needed Pilus…. “F’” Origin for transfer… “ori. T” Cam. R I 714030 Selectable marker… “Cam. R” “Part” is a genetically-encoded, human defined function ori. T F’

“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? INFECT REPLICATE RELEASE ASSEMBLE http: //escience. ws/b 572/L 17. htm

“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? INFECT REPLICATE RELEASE ASSEMBLE http: //escience. ws/b 572/L 17. htm

Genetically programmed bi-stable switch natural context: epigenetic regulation lysogeny lysis Genes Dev. (2004)18(17): 2086– 2094. doi: 10. 1101/gad. 1226004.

Genetically programmed bi-stable switch i. GEM context: flip and hold in one state promoter RBS c. I ORF transcriptional terminator lysogeny Genes Dev. (2004)18(17): 2086– 2094. doi: 10. 1101/gad. 1226004.

“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? INFECT REPLICATE RELEASE ASSEMBLE http: //escience. ws/b 572/L 17. htm

“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? INFECT REPLICATE RELEASE ASSEMBLE http: //escience. ws/b 572/L 17. htm

Fluorescence vs bioluminescence AI-1 AI-2

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

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

One last thing about fluorescence

Genetically programmed infection i. GEM context: phage infection via conjugation What “parts” are needed Pilus…. “F’” Origin for transfer… “ori. T” Selectable marker… “Cam. R” Stable lysogen… “c. I” Moniter… “GFP” Cam. R ori. T F’

Predator cell stable lysogen that infects by conjugation + Cam. R + GFP to follow process