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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
Genetic Engineering
  • Intent of altering human genome
  • Introducing new genetic material into genome
  • Isolating genes to produce on large scale (Insulin)
recombinant dna
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
process restriction enzyme
Process – Restriction Enzyme
  • Locates recognition site (Top Strand)
  • Cuts the DNA backbone
  • Locates recognition site (Bottom Strand)
  • Cuts the DNA backbone
  • DNA separates
sticky ends or blunt ends
Sticky Ends or Blunt Ends
  • Sticky ends – zigzag cuts in strand
  • Blunt ends – straight cut across strand
putting fragments together
Putting Fragments together
  • DNA ligase – sticky ends
  • T4 DNA ligase – blunt ends
    • forms phosphodiester bonds in DNA
  • 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
  • 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
identifying bacteria clones with target genes
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)
  • 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
restriction maps
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
gel electrophoresis
Gel Electrophoresis
  • Technique used to separate fragments of DNA (PCR)
  • Used to help identify information about unknown pieces of DNA (restriction maps/forensics)
  • 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

Taq polymerase is used to put strand together

  • Isolated from bacteria that live in hot springs
genetic engineering1
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
genetic engineering2
Genetic Engineering
  • Transgenic Organism (genetically modified organism, GMO)
    • Organisms that contain one or more genes from another organism
why use genetic engineering
Why use Genetic Engineering?
  • Cost
  • Larger organisms can produce larger molecules
  • Better versions of organisms
  • 80% of Canadas Canola crop is GM
gene therapy
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
gene therapy2
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
  • 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
genetic testing
Genetic Testing
  • Determining if your baby carries a gene that is associated with an increased risk of developing a disease
  • Benefits
    • Diagnosis of chromosome abnormalities
    • Diagnosis of other defects
  • Risks
    • Discomfort for expectant mother
    • Slight risk of infection
    • Miscarriage
chorionic villus sampling
Chorionic Villus Sampling
  • Benefits
    • Diagnosis of chromosome abnormalities
    • Diagnosis of certain genetic defects
  • Risks
    • Miscarriage
    • Infection
    • Newborn having limb defects
cord blood banking
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