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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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Presentation Transcript
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Summer 2008 Workshop
  • in Biology and Multimedia
  • for High School Teachers
ap biology lab 6 genetic engineering via bacterial transformation

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

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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
slide4

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:

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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

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STARTING MATERIALS

E. coli cells

  • sensitive to antibiotics
  • can’t glow
  • competent - able to be transformed

Bacterial chromosome

slide7

AmpR

Ara

promoter

STARTING MATERIALS

  • Plasmid containing:
  • Ampicillin resistance gene (always expressed)
  • Ara promoter - turned on in the presence of arabinose
slide8

GFP gene

STARTING MATERIALS

Jellyfish DNA

GFP = Green Fluorescent Protein

glows under UV light

slide9

AmpR

Ara

GFP

STARTING MATERIALS

E. coli cells

Plasmid

Jellyfish DNA

slide10

AmpR

GROW ON AN AGAR PLATE

GFP

Ara

… that can GLOW!

END RESULT

Recombinant Bacteria…

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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

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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.