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Transcript
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 Remove Plasmid DNA 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?
