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)
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

9. 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)

10. 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

11. Cloning Insulin

12. 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

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

14. PCR Polymerase Chain Reaction Process
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

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

16. 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

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

18. Knockout Mice

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

20. 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

21. Gene Therapy

22. Gene Therapy Germ-line gene therapy Somatic 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

23. Biotechnology Artificial Insemination In Vitro Fertilization
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

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

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

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

27. Embryonic Stem Cells – differentiate into any kind of cell
Stem Cell – an undifferentiated cell that can divide to form specialized cells
Embryonic Stem Cells – differentiate into any kind of cell
Muscle Cell
Skin Cell
Nerve Cell
Heart Cell
Tissue Stem Cells (Adult Stem Cells) – exist within specialized tissue, differentiate into certain types of cell s
Bone Marrow
Stem Cells

28. 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