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Warm-up

Warm-up. To which tube(s) did we add bacteria? To which tube(s) did we add plasmid? What two genes were located on the plasmid? Why are both of these genes important for our experiment?. Questions about procedure.

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Warm-up

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  1. Warm-up • To which tube(s) did we add bacteria? • To which tube(s) did we add plasmid? • What two genes were located on the plasmid? • Why are both of these genes important for our experiment?

  2. Questions about procedure • Why did we place the bacteria in hot water (42 degrees Celsius) for 50 seconds?

  3. Questions about procedure • What is meant by a “control plate”? • What purpose does a control serve?

  4. Questions about procedure • What is the purpose of the ampicilin in the growth media?

  5. Prediction • On which plates would you expect to find bacteria most like the original non-transformed E. coli colonies that you started with? • Explain your prediction.

  6. Prediction • If there are any genetically transformed bacterial cells, on which plate(s) would they most likely be located? • Explain your prediction

  7. Data Table • You will need to create a data table in your lab notebook. This table should include the following information for all four plates: • Number of Colonies on Day One • Number of Colonies on Day Two • Number of Glowing Colonies on Day Two

  8. Add arabinose to the environment of the bacteria in a controlled manner Bacteria with the gene for the green fluorescent protein will start making the protein only in the presence of arabinose. We say that the gene is “turned on” with arabinose and “turned off” in the absence of arabinose.

  9. arabinose solution 3% (0.3 g/10 ml) water

  10. Pre-cut filter paper Pre-cut filter paper arabinose solution 3% (0.3 g/10 ml) water

  11. Pre-cut filter paper Pre-cut filter paper arabinose solution 3% (0.3 g/10 ml) water Filter paper soaked in arabinose Filter paper soaked in water NOTE: Use different shapes for easy identification of which paper is soaked in arabinose and which paper is the control

  12. Experimental setup Filter paper soaked in arabinose Filter paper soaked in water LB agar, no ampicillin LB agar, + ampicillin + plasmid + plasmid no plasmid no plasmid

  13. Conclusion • Cite evidence from the bacterial transformation experiment to support the following statements:

  14. Statement 1 • The antibiotic ampicillin kills normal E. coli bacteria. • Write it down and then cite evidence from the bacterial transformation experiment to support it.

  15. Statement 2 • Only a small percentage of bacteria exposed to plasmid DNA take in the DNA during the heat shock procedure. • Write it down and then cite evidence from the bacterial transformation experiment to support it.

  16. Statement 3 • An organism’s DNA influences which proteins it makes. • Write it down and then cite evidence from the bacterial transformation experiment to support it.

  17. Next day

  18. Draw this in your lab book and label it as “Conditions” Conditions: Draw what you see on each plate. Where there are colonies, indicate which are green and which are not. Be sure to record data in your data table as well. + pGlo,+AMP +pGlo, -AMP -pGlo, +AMP -pGlo, -AMP

  19. Experimental setup Filter paper soaked in arabinose Filter paper soaked in water LB agar, no ampicillin LB agar, + ampicillin + plasmid + plasmid no plasmid no plasmid

  20. EXPECTED RESULTS Filter paper soaked in arabinose Filter paper soaked in water LB agar, no ampicillin LB agar, + ampicillin + plasmid + plasmid no plasmid no plasmid

  21. Statement 4 • An organism’s environment influences which proteins it makes. • Write it down and then cite evidence from the bacterial transformation experiment to support it.

  22. Statement 5 • One way organisms respond to their environment is by changing whether a protein is made or not. In other words, they respond to their environment sometimes by turning genes “on” or turning genes “off.” • Write it down and then cite evidence from the bacterial transformation experiment to support it.

  23. Discussion • Include how reliable you feel the results are and any errors that could have changed them. Suggest improvements to the lab to increase the validity of the results

  24. Final Assessment • In your own words, explain the relationship between these terms: • DNA • Protein • Environment • Gene • Trait

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