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Transformation Lab. Engineering a Bacteria to Glow Green. Goal:. To get E. coli to take in a plasmid that contains the GFP gene and produce green fluorescence protein when given the right environmental signal. Plasmid. Contains : Origin of Replication Antibiotic Resistance

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Transformation lab

Transformation Lab

Engineering a Bacteria to Glow Green


Goal:

  • To get E. coli to take in a plasmid that contains the GFP gene and produce green fluorescence protein when given the right environmental signal.


Plasmid
Plasmid

  • Contains:

    • Origin of Replication

    • Antibiotic Resistance

    • Restriction site to enter gene

    • Operon to control turning on/off of gene


At the biotech company
At the biotech company…

  • Restriction enzyme

  • DNA ligase


What you will do
What you will do…

  • Heat shock

  • Culture bacteria in LB

  • Plate bacteria onto 4 plates:

    • LB

    • LB/amp

    • LB/amp

    • LB/amp/ara


Will we see glowing bacteria on all our plates
Will we see glowing bacteria on all our plates?

  • Operon: Control switch that turns on/off conversion of DNARNAProtein

  • Arabinose operon: turns on the production of GFP when arabinose (sugar) is present as a signal.


Visual hypothesis what are we adding to each plate what should happen to each plate
Visual Hypothesis:What are we adding to each plate?What should happen to each plate?


We ll analyze our data tomorrow
We’ll analyze our data tomorrow!


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