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Biotechnology

Biotechnology. DNA Technology. DNA technology is the chemical manipulation of the genotypes and resulting phenotypes of organisms such that living organisms are modified Alternatively, no-longer-living organisms or their no-longer-living parts may be analyzed chemically at the level of genotype

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Biotechnology

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  1. Biotechnology

  2. DNA Technology • DNA technology is the chemical manipulation of the genotypes and resulting phenotypes of organisms such that living organisms are modified • Alternatively, no-longer-living organisms or their no-longer-living parts may be analyzed chemically at the level of genotype • DNA technology has revolutionized how scientists study the genetics, biochemistry, even the ecology and evolutionary biology of organisms • Genetic engineering is the artificial manipulation of the genetic material of organisms, including the creation of novel genetic material (i.e., novel nucleotide sequences) • Biotechnology is the development of novel biological products, indeed whole industries are now devoted to the production and analysis of biological materials

  3. Cloning Step in Overview

  4. Utilizing Cloned Genes

  5. Restriction Enzymes

  6. 5’…GAATTC…3’ 3’…CTTAAG…5’ 5’…G -OHP-AATTC…3’ 3’…CTTAA -PHO-G…5’ Restriction Endonucleases • A Restriction Endonucleases will cut both strands of a DNA duplex at a specific place • These “places” need not be directly opposite: 5’…GAATTC…3’ 3’…CTTAAG…5’ 5’…G -OHP-AATTC…3’ 3’…CTTAA -PHO-G…5’ • Note that the above enzyme is EcoRI, the first restriction endonuclease characterized

  7. Sticky Ends

  8. Most R.E. RecognitionSequences are Palindromes G^AATT-C C-TTAA^G G^GATC-C C-CTAG^G Nodeba Bob Abedon A^GATC-C T-CTAG^G GC^GGCC-GC CG-CCGG^CG

  9. More RE Enzymes

  10. DNA Ligase Upon ligation we now have recombinant DNA

  11. Ligation

  12. Cloning using Restriction Enzymes

  13. Transformation Note that plasmid is vector that carries DNA into recipient cells via transformation Other vectors include viruses (transduction) as well as otherwise inert projectiles

  14. Gene Therapy One example of this might be done… is essentially cloning into animals

  15. Recombinant Animals Alternatively, DNA can be cloned directly into the germ line

  16. Injecting DNA into Egg

  17. Cloning into Plants Many plants are easily cloned from individual body cells

  18. Selection for Specific Clones

  19. Polymerase Chain Reaction Cloning allows the amplification of genotype (DNA) as well as phenotype (proteins) If all you really need is the DNA, then PCR is an easy way to amplify DNA without cloning

  20. PCR DNA Amplification

  21. PCR DNA Amplification

  22. Gel Electrophoresis

  23. Loading Gel

  24. Loaded Gel

  25. Run Gel

  26. 2-D Protein Electrophoresis

  27. 2-D Protein Electrophoresis Visualized proteins

  28. Restriction Fragment Analysis No need to know details of what gene was studied or specific restriction enzyme used

  29. Genome Comparisons

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