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Recombinant DNA Technology

Recombinant DNA Technology. Pre-AP Biology. Bellwork. Explain what genetic engineering means to you. Key Terminology. Recombinant Chimera Plasmid Cloning Vector Ligate / DNA Ligase Transformation Transgenic. Objectives. Define recombinant DNA

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Recombinant DNA Technology

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  1. Recombinant DNA Technology Pre-AP Biology

  2. Bellwork Explain what genetic engineering means to you.

  3. Key Terminology • Recombinant • Chimera • Plasmid • Cloning Vector • Ligate / DNA Ligase • Transformation • Transgenic

  4. Objectives Define recombinant DNA Explain how bacteria are genetically engineered Create a paper model of a recombinant plasmid and identify the key components Describe examples of Biotechnology in plants and animals

  5. Overview • Recombinant DNA Technology creates new DNA molecules • made by piecing together DNA from different organisms • This recombinant DNA (rDNA) is also known as a chimera Goat boy

  6. Examples of Recombinant DNA Technology • rhInsulin • marketed by Eli, Lilly and Company • Recombinant human growth hormone • Nutropin, marketed by Genentech, Inc • Recombinant “Activase” • dissolves blockages in blood vessels • marketed by Genentech, Inc

  7. Significance of rDNA • These proteins normally are produced in very small amounts in animals • Making these proteins in large amounts in other organisms, like bacteria • Can be supplemented in individuals that are deficient in a protein, like Insulin • Can be made cheaper and safer than other methods

  8. Making a Recombinant DNA Molecule • Step 1: Isolation of human DNA encoding Insulin: • Isolate Human DNA • Cut DNA into fragments with a restriction enzyme • Identify the DNA fragment that encodes for the protein Insulin

  9. Making a Recombinant DNA Molecule • Step 2: Place DNA molecule into a cloning vector • Cloning vector is commonly a plasmid • small circular piece of DNA found in bacteria • separate from its chromosomal DNA • A plasmid is called a vector • when used to carry the desired DNA into the organism

  10. Making a Recombinant DNA Molecule (Step 2 continued) • Fragments of DNA are put into the plasmid • Cut plasmid with same restriction enzyme used to make DNA fragments • Ligate fragments into the plasmid with DNA ligase • DNA ligase “glues” the DNA fragments together

  11. Think-Pair-Share Why is the plasmid with the human insulin DNA considered a recombinant DNA molecule or chimera? 3 PROPERTY OF PIMA COUNTY JTED, 2010

  12. Transforming Bacteria with Recombinant DNA • Step 3: The recombinant DNA molecule is taken up by the Bacteria • Bacteria are said to be transformed • When they take up a foreign gene in a plasmid • e.g. when they take up the human insulin gene

  13. Transformation Process by which bacterium takes up and expresses foreignDNA Happens in nature when bacteria die and release their DNA, and other bacteria take up the DNA

  14. Think-Pair-Share IYOW define transformation of bacteria. 3 PROPERTY OF PIMA COUNTY JTED, 2010

  15. Identify the Bacteria with the Recombinant DNA molecule • Not all bacteria will be transformed • Must eliminate bacteria that do not carry the recombinant plasmid • Accomplished by growing bacteria on media containing an antibiotic • Antibiotic –chemicals that prevent the growth and survival of bacteria • How does the antibiotic work?

  16. Definition Property of CTE Joint Venture Antibiotic - “against life” substance that kills bacteria or prevents them from replicating

  17. Cloning Vector • The plasmid that is used as vector is also given an antibiotic resistance gene • The vector shown has the resistant gene to ampicillin

  18. How Bacteria Acquire Antibiotic Resistance Property of CTE Joint Venture

  19. Bacteria Take Up Cloning Vector • Bacteria that take up cloning vector will also be ampicillin resistant • In other words, transformed bacteria will not be killed when grown on agar containing ampicillin • The bacteria without the cloning vector will be killed because they do not contain the ampicillin resistance gene

  20. Think-Pair-Share You have transformed bacteria with the plasmid containing the human insulin gene. Only 10% of the bacteria take up the recombinant DNA molecule. Predict which petri plate will have more bacteria growth, the one + ampicillin or the one – ampicillin. Explain your reasoning. 3 PROPERTY OF PIMA COUNTY JTED, 2010

  21. Uses of Biotechnology in Plants • Make Transgenic Plants to • Delay ripening-ex. Flavor-Saver Tomatoes

  22. Uses of Biotechnology in Plants • Improve nutritional content • e.g. Golden Rice • Herbicide resistance • Coffee

  23. Uses of Biotechnology in Animals • Be a pharmaceutical factory • produce a large amount of an otherwise rare biological substance for medical use • Hormones • Blood clotting factors • Vaccines

  24. Q & A: Recombinant DNA • Why is it useful to create bacteria that contain a new gene? • What is the purpose of restriction enzymes in this process? • What is the purpose of DNA ligase? • Describe the 3 steps for making recombinant DNA. • Human Growth Factor is a protein. Based on what you’ve learned in this unit, what biochemical processes must take place before HGH will be produced by bacteria?

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