Transposition

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Transposition

• Barbara McClintock (1950s)
• Transposon
• Transposition
• transposon, transposase, target DNA

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Three groups of Transposons

• Insertion Sequences
• Composite transposons
• Tn10, Tn5
• Non-composite transposons
• Tn3, Tn7

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Insertion Sequences
Size 1-2 kbp
5ATGCTAC
Inverted repeat
tnp
Inverted repeat
CATCGTA-5
tnp transposase
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Models for Transposition

• Replicative transposition
• Transposon is replicated
• Donor retains a copy of transposon
• Conservative transposition
• Cut and paste

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Delivery of Transposon

• Phage-suicide vectors
• Plasmid-suicide vectors

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Applications of Transposon

• In vitro transposition
• Phage-mediated mutagenesis

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Use of phage Mu derivatives for mutagenesis
Phage Mud
Phage Mud
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Site-Specific Recombinations

• Integrases
• Resolvases
• Invertases

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Site-Specific Recombinations

• Integrases
• Promotes integration of two DNA molecules
• Recognizes specific DNA sequences
• attP in lambda phage

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Site-Specific Recombinations

• Resolvases
• Promotes recombination within the same DNA
  molecule
• Recognizes specific DNA sequence res
• Resolve the co-integrate

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