Bioengineering
Turning Genes on and off in Bacteria
If all cells have the same DNA then why aren’t all
cells the same?
Gene Expression
 Process of turning genes on and off in cells.
 Allows for differentiation.
 Gene expression is different in prokaryotes and
eukaryotes:
 Prokaryotes: operons
 Eukaryotes: DNA modification, alternative splicing,
transcription factors, RNAi
Question?
 How can we get a bacteria to express a jellyfish
gene under specific conditions?
Problem #1:
 How do we get a jellyfish gene into bacteria?
 Design a plasmid:
Plasmid Design
 Origin of replication:
 Antibiotic resistance:
 Promoter and restriction site within
operon:
Heat Shock
 Method to get bacteria to take in DNA. Bacteria take
in DNA when they are in stressful environments!
How do bacteria express
DNA that is not theirs?
 Operon: Sequence of genes that it turned on by the
presence of a specific chemical
We are going
to clone a
bacteria that
is tricked into
making green
fluorescence
protein!
pGLO lab
Introductory questions to
consider
 Where is the gene originally from?
 What does the plasmid contain?
 How will we get the plasmid into the bacteria?
 What will activate the production of GFP in bacteria?
 Google BioRAD and pGLO and find the manual and
answer these questions prior to Lab next Wednesday!
Answers
 Where is the gene originally from?
 Green Fluorescence Protein from Jellyfish
 What does the plasmid contain?
 Ampicillin resistance, Arabinose operon
 How will we get the plasmid into the bacteria?
 Heat Shock
 What will activate the production of GFP in bacteria?
 Presence of arabinose
Hypothesis:
 Visual: What will the four Petri dishes that we will
make look like?
 Written: What will cause the production of GFP in
bacteria?