1. http://www.spillerlab.com/mission/
http://www.rci.rutgers.edu/~crebb/CREBB%20Pictures/CREBB%20Web%20Photo%20A http://blogs.scientificamerican.com/guest-blog/2011/03/23/dressing-the-meat-of-
lbum/pages/GFP-EColi_jpg.htm tomorrow/

2. • GFP is isolated from the jelly
fish, Aequorea victoria
• It has been modified to enhance
is fluorescing characteristic
• You will be working with GFP
tomorrow!

3. GFP tagged Mice
Mouse Embryo
http://www4.utsouthwestern.edu/olsonlab/images.html http://www.bannination.com/comments/5047156/c1455973
Neuron
Fly
Kidney Cells
http://gfp-cdna.seebyseeing.net/
http://sciencewise.anu.edu.au/articles/tthy http://captain-nitrogen.tumblr.com/post/9257522157/this-wormy-world-indeed

4. • Take the DNA from the jelly fish and add it to the DNA of the fish
• These organisms are called transgenic organisms, because they
have genes from another organism put into their DNA
• Trans = across
• Genic = genes

5. • We are going to create a transgenic bacteria that glows in
the dark.
• We will take the GFP gene from jellyfish and put it into
E.coli bacteria
E. coli

6. • Transformation is when a cell takes in and expresses a new
piece of genetic material (DNA)
E.coli bacterial cell • E.coli is a prokaryote
bacteria
• It replicates fast
• Scientists use it as
a “cloning factory”
Bacteria DNA
New DNA piece enters E.coli cell

7. • The new DNA we will be working with is called a plasmid
• A plasmid is a circular piece of DNA, carrying one or more genes.
• The plasmid we are working with is called pGLO
New DNA piece enters E.coli cell

8. • Each of the colored regions on this plasmid, is a
gene (DNA)
• Each gene has its own function:
• araC –“turns on” GFP, in
presence of arabinose
• GFP – green fluorescent
protein (makes the
organism glow)
• bla – makes a protein that
breaks down ampicillin
• ampicillin – able to kill
bacteria

9. Green Fluorescent Protein
Beta protein breaks
down ampicillin (can
grow with ampicillin
around)
Bacteria DNA
pGLO plasmid

10. • We will be growing E.coli onto LB agar plates
• LB agar givse nutrients (food) for the bacteria
• The LB agar plates will have:
• Arabinose presence
activates GFP
• Ampicillin – which will kill
any bacteria that does not
have the pGLO plasmid
• Remember, bla gene
breaks down
ampicillin