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  1. Function of Transposase:
    • Enzyme with Endonuclease activity (nicks DNA) on both Donor and Recipient DNA
    • 1. cuts DNA causing staggered ends or sticky ends
    • 2. Inserts Donor DNA
    • 3. Has Ligase activity = Direct repeats formed in recipient DNA
  2. Cointegrate:
    DNA containing both DONOR + Recipient DNA, as a result of transposon insertion.
  3. Function of Resolvase:
    • Triggers site specific recombination between a new copy of the transposon and the original
    • SEPARATES Cointegrate (the donor + recipient DNA) into 2 SEPARATE pieces of DNA (donor + recipient)
  4. Transposons vs. Retrotransposons
    • Transposons work through DNA intermediate
    • RETROtransposons work through RNA intermediate –
    • 1. first removed from DNA
    • 2. transcribed to RNA…
    • 3. then reverse transcribed before being inserted into target DNA
    • Retrotransposons can result in Viral (long terminal repeats - LTR) or Non-viral families (Sine or LINE)
    • Enzymes used to nick transposons and retrotransposons are different
  5. Replicative vs. conservative Transposons
    • Replicative Tn3:
    • Aka COPY & paste
    • 1. Nicks in DONOR and TARGET DNA at SAME time, forming COINTEGRATE intermediate
    • 2. NO synapse made
    • 3. RESOLVASE separates donor from Target DNA
    • 4. Donor DNA is same length due to Tn3 being COPIED
    • Conservative Tn5:
    • Aka CUT & paste
    • 1. Cuts DONOR DNA first, then Target DNA later
    • 2. HAIRPIN intermediate formed
    • 3. Synapse made to align donor DNA to target
    • 4. AUTO phosphodiester bond separates donor from Target
    • 5. Donor DNA is SHORTENED by Tn5
Card Set:
2012-04-28 17:28:47

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