Biobricked High Efficiency Recombination System - PowerPoint PPT Presentation

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Biobricked High Efficiency Recombination System

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Beta: single stranded DNA binding protein, mediate recombination ... High Efficiency: 1/10. Goal: Construction of more efficient recombination system through ... – PowerPoint PPT presentation

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Title: Biobricked High Efficiency Recombination System


1
Biobricked High Efficiency Recombination System
2
Homologous Recombination
Low Efficiency lt1/104
3
Site-specific SSoligo-mediated Recombination
High Efficiency 1/10
Exo 5-3 exonuclease Beta single stranded DNA
binding protein, mediate recombination Gam
inhibits RecBCD and SbcCD, linear dsDNA degrading
nucleases
4
Goal
  • Construction of more efficient recombination
    system through mutagenesis

DL Court, 2006
Method Error prone PCR, oligo-mediated
mutagenesis
5
Advantages
  • Easy readout selection on antibiotics
  • Currently no biobricks for improved homologous
    recombination system
  • Potential for combinatorial synthesis of
    biobricks via HR
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