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Biobricked High Efficiency Recombination System

Biobricked High Efficiency Recombination System. Homologous Recombination. Low Efficiency: <1/10 4. Site-specific SSoligo-mediated Recombination. High Efficiency: 1/10. Exo: 5’-3’ exonuclease Beta: single stranded DNA binding protein, mediate recombination

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Biobricked High Efficiency Recombination System

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  1. Biobricked High Efficiency Recombination System

  2. Homologous Recombination Low Efficiency: <1/104

  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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