2.  Intent of altering human genome  Introducing new genetic material into genome  Insulin

3.  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

4.  Locates recognition site (Top Strand)  Cuts the DNA backbone  Locates recognition site (Bottom Strand)  Cuts the DNA backbone  DNA separates

5.  Sticky ends – zigzag cuts in strand  Blunt ends – straight cut across strand

6.  DNA ligase – sticky ends  T4 DNIA ligase – blunt ends › forms phosphodiester bonds in DNA

7.  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

8.  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

10.  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

11.  Cells that receive foreign DNA  Bacterial cells sometimes will not take up a plasmid  Bacteria are placed in ice water bath containing CaCl ₂  Solution is heated and cooled repeatedly disrupting plasma membrane of bacteria allowing plasmid to enter  Solution is kept at 37 ⁰ C to stabilize and grow

12.  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)

13.  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.  Technique used to separate fragments of DNA (PCR)

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

18.  Cost  Larger organisms can produce larger molecules  Better versions of organisms  80% of Canadas Canola crop is GM

21.  Techniques used to replace, remove or alter a defective gene before symptoms are expressed

22.  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

23.  To detect any mutation in DNA that is associated with a genetic disorder (huntingtons disease)  Aminocentesis › Prenatal screening

24.  Source of hematopoietic (blood forming) stem cells  Can be used to treat and cure disease and other conditions