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