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Biotechnology

Biotechnology . Chapter 13. Recombinant DNA . Tools used Restriction enzymes Gel electrophoresis DNA ligase. Restriction enzymes. Cut double stranded DNA into smaller fragments restriction sites Sticky ends Blunt ends . Gel electrophoresis. Separate and purify DNA fragments

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Biotechnology

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  1. Biotechnology Chapter 13

  2. Recombinant DNA • Tools used • Restriction enzymes • Gel electrophoresis • DNA ligase

  3. Restriction enzymes • Cut double stranded DNA into smaller fragments • restriction sites • Sticky ends • Blunt ends

  4. Gel electrophoresis • Separate and purify DNA fragments • After cutting the DNA by restriction enzymes • Placed in an electrical field • Phosphates are negatively charged so…

  5. This causes DNA to move to the positive side of the cell • Smaller molecules can move faster than larger one • This causes DNA to separate into fragments So…

  6. We get this information • The number of fragments • The size of the fragments • The relative abundance of a fragment

  7. DNA ligase • Makes phosphodiester bonds between DNA fragments. • Allows recombinant DNA You will do this in lab

  8. Cloning • Producing many identical copies of a particular DNA sequence • Transgenic cells are contain recombinant DNA

  9. Model organisms for recombinant DNA • Bacteria • Yeast • Plant cells • Cultured animal cells

  10. Vectors for recombinant DNA • Plasmids • Viruses

  11. Genomic Libraries • Collection of DNA fragments that together comprise the genome of the organism. • cDNA – complementary DNA can be found in a cDNA library • Only genes from a particular tissue (snapshot)

  12. Block translation • Antisense RNA • Used in some drugs for cancer treatment

  13. Application • Expression vectors have all the appropriate characteristics for expression • Inducible promoter • Tissue specific promoter • Signal sequences

  14. Medical products • Insulin • Human growth hormone

  15. Breeding methods • The ability to identify specific genes • Ability to introduce any gene • Ability to generate new organisms quickly

  16. Transgenic crops • Improved nutritional value • Adapted to environment

  17. Concerns • Unnatural • Safety • Enviornmental effects

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