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  • When I say GFP what does that mean?Which one is the order name? and which is the species name?How do you know?
  • Write down transgenic organisms…
  • What does transgenic mean again?
  • Bacteria can often take in floating pieces of DNA, and once inside the bacteria, the cell can often start to express the new traits from the new piece of DNA.
  • The e.coli bacteria now has all the traits of the plasmid

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  • 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/
  • • 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!
  • GFP tagged Mice Mouse Embryohttp://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 View slide
  • • 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 View slide
  • • 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) 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
  • • 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 pGLONew 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
  • Green Fluorescent Protein Beta protein breaks down ampicillin (can grow with ampicillin around)Bacteria DNA pGLO plasmid
  • • 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