Recombineering

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Recombineering is a method where E-coli undergoes recombination between linear DNA, introduced through electroporation, with circularized DNA already present within the cell.


http://web.ncifcrf.gov/research/brb/recombineeringInformation.aspx


λ phage exo, bet, and gam genes are integrated into E-coli to demonstrate phage promoted homologous recombination. Murphy 1998 λ Red-Promoted Gene Replacement


To acheive control over phage recombination gene activity, temperature sensitive λ repressor (cI857) was introduced. Yu 2000 An efficient recombination system for chromosome engineering in Escherichia coli


Court Lab.

This process is then adapted to E-coli strain DH10B, a Bacteria Artificial Chromosome (BAC)strain. This results in the original recombineering E-coli strain designated DY380 as well as other versions modified to include different selection mechanisms. Lee 2001. A highly efficient Escherichia coli-based chromosome engineering system adapted for recombinogenic targeting and subcloning of BAC DNA

For additional information on protocols, strains, and recombineering agents being developed by the Court lab at NCI-Frederick please check their website: http://redrecombineering.ncifcrf.gov/


Church Lab.

Methods for generating precise deletions and insertions in the genome of wild-type Escherichia coli: application to open reading frame characterization-



All about the λ phage recombination proteins, exonuclease, β, and γ!!


An Exonuclease Induced by phage λ. Little 1967

Roles of Exonuclease and β protein of phage λ in recombination. Carter 1971


β protein of phage λ promotes renaturation of DNA. Kmiec 1981

Pairing activities of β protein in phage λ. Muniyappa 1986


How γ protein specified by phage λ affects recBC enzyme in E. Coli. Karu 1975

γ protein of phage λ inhibits recombination activites of RecBCD enzyme. Murphy 1991

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