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Summer 2008 Workshop in Biology and Multimedia for High School Teachers

Summer 2008 Workshop in Biology and Multimedia for High School Teachers. AP Biology Lab 6: Genetic Engineering via Bacterial Transformation. Making E. coli glow like jellyfish. Amy Dickson, Prospect Hill Academy Charter School All images by Christine Rodriguez and Amy Dickson.

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Summer 2008 Workshop in Biology and Multimedia for High School Teachers

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  1. Summer 2008 Workshop • in Biology and Multimedia • for High School Teachers

  2. AP Biology Lab 6:Genetic EngineeringviaBacterial Transformation Making E. coli glow like jellyfish Amy Dickson, Prospect Hill Academy Charter School All images by Christine Rodriguez and Amy Dickson

  3. WHY SHOULD WE DO THIS? Genetic Engineering is now widely used: • Bacteria that produce human insulin • Corn that produces insecticide • Rice that produces extra vitamin A • Goats that produce spider silk

  4. DNA RNA Protein Trait Green Fluorescent Protein • GFP Gene • found in jellyfish • engineered into bacteria GLOWING CELLS WHY SHOULD WE DO THIS? To SEE the Central Dogma in action:

  5. QUICK REVIEW Promoter - Plasmid - Transformation - an “on/off” switch for a gene a small, circular piece of bacterial DNA that is not part of the chromosome a process in which bacteria take up DNA from their environment - can be triggered by electric shock or heat shock

  6. STARTING MATERIALS E. coli cells • sensitive to antibiotics • can’t glow • competent - able to be transformed Bacterial chromosome

  7. AmpR Ara promoter STARTING MATERIALS • Plasmid containing: • Ampicillin resistance gene (always expressed) • Ara promoter - turned on in the presence of arabinose

  8. GFP gene STARTING MATERIALS Jellyfish DNA GFP = Green Fluorescent Protein glows under UV light

  9. AmpR Ara GFP STARTING MATERIALS E. coli cells Plasmid Jellyfish DNA

  10. AmpR GROW ON AN AGAR PLATE GFP Ara … that can GLOW! END RESULT Recombinant Bacteria…

  11. makes all transformed bacteria resistant to ampicillin controls GFP gene expression only turned on in the presence of arabinose HOWEVER… things are actually a bit more complex. AmpR pGLO plasmid GFP Ara promoter

  12. pGLO YOUR TASK: Design an experimental procedure for genetically engineering glowing bacteria. Goals to consider: #1 - Make recombinant bacteria #2 - Select for only the recombinant bacteria #3 - Make the recombinant bacteria glow #4 - Establish a control for your experiment to demonstrate that it’s the plasmid that causes ampicillin resistance and the ability to glow.

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