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Hoofdstuk 20 2008 deel 1

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Hoofdstuk 20 2008 deel 1 Hoofdstuk 20 2008 deel 1 Presentation Transcript

  • Chapter 20 DNA technology
  • Outline of this chapter on techniques
    • DNA analysis
      • Restriction enzymes
        • Recombinant DNA
      • Libraries
      • Making cDNA
      • Hybridization
      • PCR, RT-PCR
        • Reverse transcriptase-polymerase chain reaction
      • Electrophoresis
      • Southern blotting
      • DNA sequencing
      • Micro-arrays
      • Cloning
        • Plants
        • Bacteria
        • Animals
        • Use of stem cells
    • Practical applications
      • SNPs
        • Single nucleotide polymorphism
      • RFLP, diagnosis of disease
      • Gene therapy
      • Transgenic plants
    • DNA cloning
      • Origin of gene
      • Use of a plasmid
      • Cloning gene into plasmid
      • Selection of recombinant bacteria
      • Multiplication of bacteria
    Computermodule supports study of this chapter
  • “ Wat moet ik leren van dit hoofdstuk?”
    • Te bestuderen:
      • Afhankelijk van de nog beschikbare tijd, maar in ieder geval:
      • Kloneren van een gen in een plasmide
        • Ook practicum!
      • PCR: polymerase chain reaction
      • DNA sequencing
  • 2 Overview of the use of a bacterial plasmid in cloning genes
  • 2 Overview of the use of a bacterial plasmid in cloning genes
  • 2 Overview of the use of a bacterial plasmid in cloning genes Transformation
  • 2 Overview of the use of a bacterial plasmid in cloning genes Transformation
  • 2 Overview of the use of a bacterial plasmid in cloning genes Transformation
  • 2 Overview of the use of a bacterial plasmid in cloning genes Transformation
  • 2 Applications of cloned genes
  • 3 Using a restriction enzyme to make recombinant DNA
  • 3 Using a restriction enzyme to make recombinant DNA
    • Palindromic sequence:
        • GAA|TTC CTT|AAG
    • Sticky ends
      • 5’ overhang
        • … .G ….CTTAA
      • 3’ overhang
        • … .GAATT ….C
    • Blunt ends
        • … GAA …CTT
    • Restriction
      • bacteria
      • modification by methylation
    • Ligation gives two combinations!!
  • 3 Using a restriction enzyme to make recombinant DNA
    • Palindromic sequence:
        • GAA|TTC CTT|AAG
    • Sticky ends
      • 5’ overhang
        • … .G ….CTTAA
      • 3’ overhang
        • … .GAATT ….C
    • Blunt ends
        • … GAA …CTT
    • Restriction
      • bacteria
      • modification by methylation
    • Ligation gives two combinations!!
  • 3 Using a restriction enzyme to make recombinant DNA
    • Palindromic sequence:
        • GAA|TTC CTT|AAG
    • Sticky ends
      • 5’ overhang
        • … .G ….CTTAA
      • 3’ overhang
        • … .GAATT ….C
    • Blunt ends
        • … GAA …CTT
    • Restriction
      • bacteria
      • modification by methylation
    • Ligation gives two combinations!!
  • 3 Using a restriction enzyme to make recombinant DNA
    • Palindromic sequence:
        • GAA|TTC CTT|AAG
    • Sticky ends
      • 5’ overhang
        • … .G ….CTTAA
      • 3’ overhang
        • … .GAATT ….C
    • Blunt ends
        • … GAA …CTT
    • Restriction
      • bacteria
      • modification by methylation
    • Ligation gives two combinations!!
  • 3 Using a restriction enzyme to make recombinant DNA
    • Palindromic sequence:
        • GAA|TTC CTT|AAG
    • Sticky ends
      • 5’ overhang
        • … .G ….CTTAA
      • 3’ overhang
        • … .GAATT ….C
    • Blunt ends
        • … GAA …CTT
    • Restriction
      • bacteria
      • modification by methylation
    • Ligation gives two combinations!!
  • 4 & 5 Cloning a bird gene in a plasmid (COO + practicals!)
    • Cloning vector
      • A plasmid
      • Contains
        • amp-gene: resistance against ampicillin
          • a penicillin
      • lacZ gene (ß-galactosidase)
        • Converts X-gal into a blue product
    • Screen for amp R , lacZ -
    • One colony is a clone from one bacterium
  • 4 & 5 Cloning a bird gene in a plasmid (COO + practicals!)
    • Cloning vector
      • A plasmid
      • Contains
        • amp-gene: resistance against ampicillin
          • a penicillin
      • lacZ gene (ß-galactosidase)
        • Converts X-gal into a blue product
    • Screen for amp R , lacZ -
    • One colony is a clone from one bacterium
    Filmpje: cloning A library of hummingbird genes