- λ red-mediated gene replacement — Datsenko and Wanner method, using plasmids
- Court lab λRed temperature sensitive system — Court method using temperature sensitive plasmids (pSIM5)
Recombineering is a method where E-coli undergoes recombination between linear DNA, introduced through electroporation, with circularized DNA already present within the cell.
[math]\lambda[/math] phage exo, bet, and gam genes are integrated into E-coli to demonstrate phage promoted homologous recombination. Murphy 1998 [math]\lambda[/math] Red-Promoted Gene Replacement
To acheive control over phage recombination gene activity, temperature sensitive [math]\lambda[/math] repressor (cI857) was introduced. Yu 2000 An efficient recombination system for chromosome engineering in Escherichia coli
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/
Methods for generating precise deletions and insertions in the genome of wild-type Escherichia coli: application to open reading frame characterization-
All about the [math]\lambda[/math] phage recombination proteins, exonuclease, [math]\beta[/math], and [math]\gamma[/math]!!
An Exonuclease Induced by phage [math]\lambda[/math]. Little 1967
Roles of Exonuclease and [math]\beta[/math] protein of phage [math]\lambda[/math] in recombination. Carter 1971
[math]\beta[/math] protein of phage [math]\lambda[/math] promotes renaturation of DNA. Kmiec 1981
Pairing activities of [math]\beta[/math] protein in phage [math]\lambda[/math]. Muniyappa 1986
How [math]\gamma[/math] protein specified by phage [math]\lambda[/math] affects recBC enzyme in E. Coli. Karu 1975
[math]\gamma[/math] protein of phage [math]\lambda[/math] inhibits recombination activites of RecBCD enzyme. Murphy 1991