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Genetic Engineering

Genetic Engineering. Intent of altering human genome Introducing new genetic material into genome Isolating genes to produce on large scale ( Insulin). Recombinant DNA. DNA that contains genes of two species How? Restriction enzymes – cut out desired gene

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Genetic Engineering

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  1. Genetic Engineering • Intent of altering human genome • Introducing new genetic material into genome • Isolating genes to produce on large scale (Insulin)

  2. Recombinant DNA • DNA that contains genes of two species • How? • Restriction enzymes – cut out desired gene • Occur naturally in prokaryotic cells • Recognize specific recognition sites – 4 to 8 base pairs • Recognition sites are palindromes • Cuts gene (digests) in one direction only • Creates restriction fragments

  3. Process – Restriction Enzyme • Locates recognition site (Top Strand) • Cuts the DNA backbone • Locates recognition site (Bottom Strand) • Cuts the DNA backbone • DNA separates

  4. Sticky Ends or Blunt Ends • Sticky ends – zigzag cuts in strand • Blunt ends – straight cut across strand

  5. Putting Fragments together • DNA ligase – sticky ends • T4 DNA ligase – blunt ends • forms phosphodiester bonds in DNA

  6. Plasmids • Small circular pieces of DNA found in bacteria • Used as vectors for recombinant DNA (artificial) • Restriction enzymes used to isolate specific gene are used to cut plasmids

  7. Process • Plasmids and DNA fragments are placed in same solution • Anneal • DNA ligase is used to form phosphdiester bond • Recombinant DNA introduced into host cell • DNA is cloned

  8. Identifying Bacteria Clones with Target Genes • Hybridization – identify cells that contain recombinant DNA • Identified using a hybridization probe – short single stranded complementary DNA molecule • Once identified bacteria can be grown in huge quantities (commercial use)

  9. Transformation • Cells that receive foreign DNA • Bacterial cells sometimes will not take up a plasmid • Bacteria are placed in ice water bath containing CaCl₂ • Solution is heated and cooled repeatedly disrupting plasma membrane of bacteria allowing plasmid to enter • Solution is kept at 37⁰C to stabilize and grow

  10. Cloning Insulin

  11. Restriction Maps • Diagrams that show all recognition sites on a specific plasmid and distances in base pairs • Shows which restriction enzyme should be used • Allow scientists to determine which plasmids will work the best for cloning experiments

  12. Gel Electrophoresis • Technique used to separate fragments of DNA (PCR) • Used to help identify information about unknown pieces of DNA (restriction maps/forensics)

  13. PCR • Polymerase Chain Reaction • Increase number of DNA copies from a single biological sample in a few hours • Only specific regions of a chromosome are replicated • Process • Denaturation • Annealing • Elongation

  14. Taq polymerase is used to put strand together • Isolated from bacteria that live in hot springs

  15. Genetic Engineering • Biopharming • Pharmaceutical products produced on large scale • Organisms are genetically engineered to produce a specific protein • Ability to make new protein is passed on to offspring

  16. Genetic Engineering • Transgenic Organism (genetically modified organism, GMO) • Organisms that contain one or more genes from another organism

  17. Knockout Mice

  18. Why use Genetic Engineering? • Cost • Larger organisms can produce larger molecules • Better versions of organisms • 80% of Canadas Canola crop is GM

  19. Gene Therapy • Techniques used to replace, remove or alter a defective gene before symptoms are expressed • Insert healthy genes into cells of tissues that are affected by a disorder

  20. Gene Therapy

  21. Gene Therapy • Germ-line gene therapy • Genes introduced in sperm or egg cells • Passed on to future generations • Somatic gene therapy • Genes introduced into body cells (not sperm or egg) • Will not be passed on

  22. Biotechnology • Artificial Insemination • Sperm is collected and concentrated before being placed in woman’s uterus • In Vitro Fertilization • Eggs from mom are retrieved and combined with sperm in petri dish • After fertilization, zygote is placed in woman’s uterus

  23. Genetic Testing • Determining if your baby carries a gene that is associated with an increased risk of developing a disease

  24. Aminocentesis • Benefits • Diagnosis of chromosome abnormalities • Diagnosis of other defects • Risks • Discomfort for expectant mother • Slight risk of infection • Miscarriage

  25. Chorionic Villus Sampling • Benefits • Diagnosis of chromosome abnormalities • Diagnosis of certain genetic defects • Risks • Miscarriage • Infection • Newborn having limb defects

  26. Stem Cells

  27. Cord Blood Banking • Source of hematopoietic (blood forming) stem cells • Blood (stem cells) found in umbilical cord after birth • Similar to tissue stem cells • Develop into various blood cells • These stem cells could be banked (stored) • Used to treat variety of diseases

  28. http://www.youtube.com/watch?v=8JTw2RpDo9o&feature=related http://www.youtube.com/watch?v=VOXJaIvRltM http://www.youtube.com/watch?v=czzCmgr0IgQ&feature=related http://www.youtube.com/watch?v=VOXJaIvRltM http://www.youtube.com/watch?v=qiQfGBOoMPM&feature=fvst http://www.youtube.com/watch?v=w9wIxe4WGFg&feature=related http://www.youtube.com/watch?v=L-Rj_rvBLZ0&feature=related http://www.youtube.com/watch?v=-FaGf0h20Vw

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