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Introduction to: Bacteriophage Genetics

Introduction to: Bacteriophage Genetics. http://hiv.sourceforge.net/launch/. Why  ?. Bacteriophages are the bacterial equivalent to Viruses – we have learned a lot on Viral mechanisms from phages. Handling of phages is similar to handling of Viruses – just simpler!

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Introduction to: Bacteriophage Genetics

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  1. Introduction to: Bacteriophage Genetics http://hiv.sourceforge.net/launch/

  2. Why ? • Bacteriophages are the bacterial equivalent to Viruses – we have learned a lot on Viral mechanisms from phages. • Handling of phages is similar to handling of Viruses – just simpler! • Viruses causes immediately life threatening diseases like SARS and EBOLA, chronically diseases like AIDS and cancer and everyday diseases like common cold and influenza. • Studies on phages was the single most important contributions to development of the “Molecular Biology”.

  3. Reports • Deadlines: • 2 weeks after completion: Tuesday April 8’th 2008. • No late reports will be accepted! • Questions: Tuesday April 1’st 13.15 – 15.00 in my office in 16.2 • Reports are delivered in my letter box: Ole Skovgaard in 18.1 • I must have corrected all reports before May 16’th 2008 - and you turn in corrected reports in due time that I can fulfill this!

  4. Viral life cycle, simple version: Picture from Brock 10’th Ed.

  5. Comparison of virus with cells: Phage receptors: lamB (malB) T6 tsx T1 tonB • Cell: Phage/Virus: • Protein Protein • DNA DNA or RNA • RNA • Lipider (Lipider) • Cellwall • components • - • Phages / Viruses feed on cells!

  6. Examples of different Coliphages R.G. Glass: Gene Function, Crohm Helm 1982 London

  7. Plating Plating: Pictures from Brock 10’th Ed.

  8. I: Genetic Recombination • Genetic distance: Distance % • Cotransduktion % • Centimorgan • Minutes • Bp

  9. II: Genetic complementation Figure 6.4: Cis-trans test. Drying of plates before spot tests: 10' @ 37 °C with fan.

  10. III: Induction of lysogenic bacteria Figure 6.5: Lytic and lysogenic cycle (Modified from: A. Lwoff: "Lysogeny" Bacterial Rev. 17:269-337 (1953) .

  11. III: Induction of lysogenic bacteria The cI857 protein is temperature sensitive: Active @ 30°C and inactive 42°C PE: Establishing lysogeny; requires cII PM: Maintains lysogeny, autoregulated by cI PL and PR: major early promoters of lysis

  12. IV: Restriction / Modification

  13. How to work efficiently in the lab: • Object: • Phage techniques and classical experiments. • Management of several ongoing parallel experiments: • Before starting an experiment: Study the manual. Compare text and flow diagrams. Make sure to understand the reason for each step. • Before leaving an experiment: Make sure you have the following for the next step(s): • Knowledge: what to do and why. • Materials: what you need and ready to use. • And then you can do the other experiment or have a break!

  14. Notes for the statistics: You have this formula: (3) Of which type is that?

  15. Notes for the statistics: For this formula: The standard deviation, s, is calculated as:

  16. Notes for the statistics: How can you estimate s? Theoretical: for a Poisson distribution s is given by: Estimate from the observed standard error, SE:

  17. Notes: • Practical: • Phages and cultures are place on your bench top. • Phages and cultures are diluted and ready for use. • Inspection of plates: Tuesday @ 13:00. • Questions for the report: • While reading the plates. • Tuesday, April 1’st 13.15 – 15.00 in my office in 16.2. • Safety: • Chloroform & chloroform waste stays in the fume hood. • Other?

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