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?