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?