http://www.rci.rutgers.edu/~crebb/CREBB%20Pictures/CREBB%20Web%20Photo%20Album/pages/GFP-EColi_jpg.htm

http://blogs.scientificamerican.com/guest-blog/2011/03/23/dressing-the-meat-of-

tomorrow/

http://www.spillerlab.com/mission/

• 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!

http://www4.utsouthwestern.edu/olsonlab/images.html

Mouse Embryo

GFP tagged Mice

http://www.bannination.com/comments/5047156/c1455973

Kidney Cells

http://sciencewise.anu.edu.au/articles/tthy

Neuron

http://captain-nitrogen.tumblr.com/post/9257522157/this-wormy-world-indeed

http://gfp-cdna.seebyseeing.net/

Fly

• 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

• 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

• Transformation is when a cell takes in and expresses a new piece of genetic material (DNA)

New DNA piece enters E.coli cell

E.coli bacterial cell • E.coli is a prokaryote bacteria

• It replicates fast• Scientists use it as

a “cloning factory”

Bacteria DNA

• 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

• 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

pGLO plasmid

Green Fluorescent Protein

Beta protein breaks down ampicillin (can grow with ampicillin around)

Bacteria DNA

• 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