1. What is Willy Wonka famous for?

2. Can you remember any of the imaginative technologies Willy Wonka created?

3. Who worked for him? Oompa Loompas They’re reaching retirement age!

4. 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.

5. 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

6. 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!

7. I. Genetically Engineer a Plasmid 5 Steps Step 1: Remove DNA from the Choco Plant cell Remove DNA

8. Step 2: Remove plasmid DNA from bacteria cell Bacteria Plasmid DNA Remove Plasmid DNA

9. 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).

10. Step 4: “Glue” the sticky ends of the gene to the sticky ends of the plasmid with another enzyme called ligase. Ligase

11. Step 5: Put recombinant plasmid back into bacteria. Now this bacteria is genetically engineered! Bacteria

12. 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

13. II. Amplify Amplify the bacteria: this is making clones. Bacteria is producing Choco-M protein (and other bacterial proteins, too) Clones

14. 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.

15. 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!

16. How did you separate the two proteins? What property of the protein did you use?

17. Real-life Example: Insulin Diabetics need insulin to help regulate blood sugar. Can you design a protocol for making HUMAN INSULIN in bacteria cells?