p glo presentation
TRANSCRIPT
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