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What is Willy Wonka famous for?
Can you remember any of the imaginative
technologies Willy Wonka created?
Who worked for him?
• Oompa Loompas• They’re reaching
retirement age!
Willy Wonka is looking into new technologies to
produce candy.• One technology
he’s working with is genetic engineering.
• Goal: Learn how genetic engineering works and apply the technology to other situations.
What is Genetic Engineering?
• Process where DNA from one species is transferred into DNA of another species.
• The new strand (with DNA from both species) is called recombinant DNA.
Bacteria
Willy Wonka’s Genetic Engineering Experiment
• He wanted to make large quantities of Choco-M, a protein that is sweet and chocolaty.
• How did he do it?• In five steps!
I. Genetically Engineer a Plasmid
• 5 Steps• Step 1: Remove DNA from the
Choco Plant cell
Remove DNA
• Step 2: Remove plasmid DNA from bacteria cell
Bacteria
Plasmid DNA
Remove Plasmid DNA
• Step 3: Cut both with a restriction enzyme.
a. Cut out Cocoa-M gene
b. Cut plasmid with the same restriction enzyme
c. These cuts make “sticky ends” (unparied bases at the ends of the strands).
• Step 4: “Glue” the sticky ends of the gene to the sticky ends of the plasmid with another enzyme called ligase.
Ligase
• Step 5: Put recombinant plasmid back into bacteria. Now this bacteria is genetically engineered!
Bacteria
Production of Choco-M
• Do people want to eat bacteria and chocolate mixed together?
• What does Willy Wonka want to do now?
• Make a lot of product• Have a pure form of chocolate
II. Amplify
• Amplify the bacteria: this is making clones.
• Bacteria is producing Choco-M protein (and other bacterial proteins, too)
Clones
III. Isolate and Purify
• Break open bacterial cells with lysozyme.
• We only want Choco-M protein.• Right now it’s mixed with other
bacterial proteins.
Imagination
• How could we quickly and efficiently separate the 2 kinds of proteins?
• Keep in mind—thousands of proteins will need to be separated every hour!
• Use tools from the big toolbox…or your imagination!
• How did you separate the two proteins?
• What property of the protein did you use?
Real-life Example: Insulin
• Diabetics need insulin to help regulate blood sugar.
• Can you design a protocol for making HUMAN INSULIN in bacteria cells?