biobricked high efficiency recombination system
Post on 04-Jan-2016
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Biobricked High Efficiency Recombination System. Homologous Recombination. Low Efficiency:
TRANSCRIPT
Biobricked High Efficiency Recombination System
Homologous Recombination
Low Efficiency: <1/104
Site-specific SSoligo-mediated Recombination
Exo: 5’-3’ exonuclease
Beta: single stranded DNA binding protein, mediate recombination
Gam: inhibits RecBCD and SbcCD, linear dsDNA degrading nucleases
High Efficiency: 1/10
Goal:
Construction of more efficient recombination system through mutagenesis
DL Court, 2006
Method: Error prone PCR, oligo-mediated mutagenesis
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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