Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard College. Summer 2008...
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Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard
College.
• Summer 2008 Workshop
• in Biology and Multimedia
• for High School Teachers
Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard
College.
AP Biology Lab 6:Genetic Engineering
viaBacterial Transformation
Making E. coli glow like jellyfish
Amy Dickson, Prospect Hill Academy Charter School
All images by Christine Rodriguez and Amy Dickson
Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard
College.
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
Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard
College.
WHY SHOULD WE DO THIS?
To SEE the Central Dogma in action:
DNA
RNA
Protein
Trait
GFP Gene
• found in jellyfish
• engineered into bacteria
Green Fluorescent Protein
GLOWING CELLS
Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard
College.
QUICK REVIEW
Promoter -
Plasmid -
Transformation - a process in which bacteria take up DNA from their environment
a small, circular piece of bacterial DNA that is not part of the chromosome
an “on/off” switch for a gene
- can be triggered by electric shock or heat shock
Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard
College.
STARTING MATERIALS
Bacterial chromosome
E. coli cells
• sensitive to antibiotics
• can’t glow
• competent - able to be transformed
Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard
College.
STARTING MATERIALS
AmpR
Ara
promoter
Plasmid containing:
• Ampicillin resistance gene (always expressed)
• Ara promoter - turned on in the presence of
arabinose
Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard
College.
STARTING MATERIALS
GFP gene
Jellyfish DNA
GFP = Green Fluorescent Protein
glows under UV light
Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard
College.
GFP
Jellyfish DNA
STARTING MATERIALS
E. coli cells
AmpR
Ara
Plasmid
Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard
College.
END RESULT
AmpR
Ara
GFP
Recombinant Bacteria…
… that can GLOW!
GROW ON AN AGAR PLATE
Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard
College.
HOWEVER…things are actually a bit more
complex.
AmpR
Arapromoter
GFP
pGLO plasmid
makes all transformed bacteria resistant to ampicillin
controls GFP gene expression
only turned on in the presence of arabinose
Life Sciences-HHMI Outreach. Copyright 2008 President and Fellows of Harvard
College.
YOUR TASK:Design an experimental procedure for genetically engineering glowing
bacteria.
pGLO
Goals to consider:
#1 - Make recombinant bacteria
#2 - Select for only the recombinant bacteria
#4 - Establish a control for your experiment to demonstrate that it’s the plasmid that causes ampicillin resistance and the ability to glow.
#3 - Make the recombinant bacteria glow