Recombinant DNA technology

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  • Recombinant DNA Technology
    • The transfer of DNA fragments from one organism, or species, to another
      • The transferred DNA can be translated within cells of new organism = universal code
    • Conversion of mRNA to complementary DNA(cDNA), using reverse transcriptase
      • Reverse transcriptase catalyses the formation of double stranded DNA from single stranded RNA
        • Produces cDNA that acts as genes from extracted mRNA
    • Using restriction enzymes to cut DNA fragments
      • Restriction endonucleases cut DNA at specific sequences, leaving sticky ends
        • Same endonucleases are used to cut 2 DNA fragments then the sticky ends will be complementary and attach
    • Creating gene with 'gene machine'
      • Desired sequence is made. Produces oligonucleotides. These join to form larger DNA fragments
    • In vitro
    • In vivo
    • Benefits
    • Issues
      • Industry          -Sometimes GM foods  can't be imported or produced        - loss of profit   - no labels, no choice
  • 1. Start with DNA sample, free nucleotides, primers and DNA polymerase
  • 2. Heat to 95C to break H bonds and separate strands.
  • 3. Cool to 55C to allow primers to bind to complementary bases on DNA
  • 4. Heated to 72C for DNA polymerase to line up free nucleotides and create a new strand.
  • 5. Repeat so more fragments are obtained
  • 1. Plasmid and isolated gene are cut with same restriction enzyme, creating complementary sticky ends and joined by DNA ligase.
  • 2. Vector with recombinant DNA must be incorporated into bacteria's DNA ( by infection)
  • - Promoter and terminator sequence is added, or desired gene is not expressed
  • 3. Gene marker ( e.g. fluorescent marker, antibiotic resistance marker or enzyme marker) is part of the plasmid. Therefore, plasmids with recombinant DNA are identified.
  • Agriculture      - Controls malnutrition, famine             -Reduces use of pesticides     - Cost effective
  • Industry          - Biological catalysts are produced in large amounts from transformed organisms
  • Medicine         - Drugs, vaccines are made quickly, in large amounts and cheaply
  • Medicine         - Could be used unethically      - Less money available for resources
  • Agriculture       - Could be resistant to herbicides      - Prone to diseases

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