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Biotechnology and Recombinant DNA

Biotechnology and Recombinant DNA. Biotechnology: The use of microorganisms, cells, or cell components to make a product Foods, antibiotics, vitamins, enzymes Recombinant DNA Technology: Insertion or modification of genes to produce desired proteins. DNA Cloning. Figure 9.1.1. DNA Cloning.

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Biotechnology and Recombinant DNA

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  1. Biotechnology and Recombinant DNA • Biotechnology: • The use of microorganisms, cells, or cell components to make a product • Foods, antibiotics, vitamins, enzymes • Recombinant DNA Technology: • Insertion or modification of genes to produce desired proteins

  2. DNA Cloning Figure 9.1.1

  3. DNA Cloning Figure 9.1.2

  4. E. coli • Used because it is easily grown and its genomics are known • Need to eliminate endotoxin from products • Cells must be lysed to get product

  5. Recombinant Therapeutics Table 9.1.1

  6. Recombinant Therapeutics Table 9.1.2

  7. Selection & Mutation • Selection: Culture a naturally-occurring microbe that produces desired product • Mutation: Mutagens cause mutations that might result in a microbe with a desirable trait • Site-directed mutagenesis: Change a specific DNA code to change a protein

  8. Restriction Enzymes • Used in Cloning • Cut specific sequences of DNA • Isolated from bacterial cells

  9. Use of Restriction Enzymes Figure 9.2

  10. Vectors • Carry new DNA to desired cell • Shuttle vectors can exist in several different species • Plasmids and viruses can be used as vectors Figure 9.3

  11. Polymerase Chain Reaction (PCR) • To make multiple copies of a piece of DNA enzymatically • Used to • Clone DNA for recombination • Add restriction sites to your gene • Sequence DNA • Diagnose genetic disease • Detect pathogens

  12. PCR Figure 9.4.1

  13. PCR Figure 9.4.2

  14. How Do Scientists Insert DNA into a Cells? • Transformation • Transduction (Ch. 12) • Electroporation Figure 9.5b

  15. DNA can be inserted into a cell by: • Microinjection • Gene gun Figure 9.6 & 7

  16. Obtaining DNA • Gene libraries are made of pieces of an entire genome stored in plasmids or phages • cDNA is made from mRNA by reverse transcriptase • Synthetic DNA is made by a DNA synthesis machine

  17. Genetic Engineering: Cloning a Gene Figure 9.11.1

  18. Genetic Engineering Figure 9.11.2

  19. Making a Gene Product: Colony Hybridization Figure 9.12.1

  20. Making a Gene Product: Colony Hybridization Figure 9.12.2

  21. Therapeutic Applications • Subunit vaccines • Nonpathogenic viruses carrying genes for pathogen's antigens as vaccines • Gene therapy to replace defective or missing genes • Human Genome Project • Nucleotides have been sequenced • Human Proteome Project may provide diagnostics and treatments

  22. Scientific Applications • Understanding of DNA • Sequencing organisms' genomes • DNA fingerprinting for identification Figure 9.16

  23. Southern Blotting Figure 9.15.1

  24. Southern Blotting Figure 9.15.2

  25. Southern Blotting Figure 9.15.3

  26. Safety Issues and Ethics • Avoid accidental release • Genetically modified crops must be safe for consumption and for the environment • Who will have access to an individual's genetic information?

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