Recombineering: Difference between revisions
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==Protocols== | |||
*[[/Lambda red-mediated gene replacement|λ red-mediated gene replacement]] — Datsenko and Wanner method, using plasmids | |||
*[[Sauer:Plasmid-born λRed recombineering using dsDNA|Court lab λRed temperature sensitive system]] — Court method using temperature sensitive plasmids (pSIM5) | |||
==Resources== | |||
Recombineering is a method where E-coli undergoes recombination between linear DNA, introduced through [[electroporation]], with circularized DNA already present within the cell. | Recombineering is a method where E-coli undergoes recombination between linear DNA, introduced through [[electroporation]], with circularized DNA already present within the cell. | ||
http:// | http://web.ncifcrf.gov/research/brb/recombineeringInformation.aspx | ||
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[http://www.pnas.org/cgi/content/abstract/97/11/5978 Yu 2000 An efficient recombination system for chromosome engineering in Escherichia coli] | [http://www.pnas.org/cgi/content/abstract/97/11/5978 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. | 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. | ||
[http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11352566 Lee 2001. A highly efficient Escherichia coli-based chromosome engineering system adapted for recombinogenic targeting and subcloning of BAC DNA] | [http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=11352566 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[http://jb.asm.org/cgi/content/abstract/179/20/6228 -] | |||
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>. [http://www.jbc.org/cgi/content/abstract/242/4/679 Little 1967] | |||
Roles of Exonuclease and <math>\beta</math> protein of phage <math>\lambda</math> in recombination. [http://www.jbc.org/cgi/content/abstract/246/8/2502 Carter 1971] | |||
<math>\beta</math> protein of phage <math>\lambda</math> promotes renaturation of DNA. [http://www.jbc.org/cgi/content/abstract/256/24/12636 Kmiec 1981] | |||
Pairing activities of <math>\beta</math> protein in phage <math>\lambda</math>. [http://www.jbc.org/cgi/content/abstract/261/16/7472 Muniyappa 1986] | |||
How <math>\gamma</math> protein specified by phage <math>\lambda</math> affects recBC enzyme in E. Coli. [http://www.jbc.org/cgi/content/abstract/250/18/7377 Karu 1975] | |||
[http://www.jbc.org/cgi/content/abstract/ | |||
<math>\gamma</math> protein of phage <math>\lambda</math> inhibits recombination activites of RecBCD enzyme. [http://jb.asm.org/cgi/gca?sendit=Get+All+Checked+Abstract%28s%29&SEARCHID=1&VOLUME=173&FIRSTPAGE=5808&FIRSTINDEX=0&hits=10&RESULTFORMAT=&gca=jb%3B173%2F18%2F5808 Murphy 1991] | |||
[http://jb.asm.org/cgi/gca?sendit=Get+All+Checked+Abstract%28s%29&SEARCHID=1&VOLUME=173&FIRSTPAGE=5808&FIRSTINDEX=0&hits=10&RESULTFORMAT=&gca=jb%3B173%2F18%2F5808 Murphy 1991] | |||
[ | [[Category:Escherichia coli]] |
Latest revision as of 09:56, 1 February 2012
Protocols
- λ red-mediated gene replacement — Datsenko and Wanner method, using plasmids
- Court lab λRed temperature sensitive system — Court method using temperature sensitive plasmids (pSIM5)
Resources
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
[math]\displaystyle{ \lambda }[/math] phage exo, bet, and gam genes are integrated into E-coli to demonstrate phage promoted homologous recombination.
Murphy 1998 [math]\displaystyle{ \lambda }[/math] Red-Promoted Gene Replacement
To acheive control over phage recombination gene activity, temperature sensitive [math]\displaystyle{ \lambda }[/math] 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 [math]\displaystyle{ \lambda }[/math] phage recombination proteins, exonuclease, [math]\displaystyle{ \beta }[/math], and [math]\displaystyle{ \gamma }[/math]!!
An Exonuclease Induced by phage [math]\displaystyle{ \lambda }[/math]. Little 1967
Roles of Exonuclease and [math]\displaystyle{ \beta }[/math] protein of phage [math]\displaystyle{ \lambda }[/math] in recombination. Carter 1971
[math]\displaystyle{ \beta }[/math] protein of phage [math]\displaystyle{ \lambda }[/math] promotes renaturation of DNA. Kmiec 1981
Pairing activities of [math]\displaystyle{ \beta }[/math] protein in phage [math]\displaystyle{ \lambda }[/math]. Muniyappa 1986
How [math]\displaystyle{ \gamma }[/math] protein specified by phage [math]\displaystyle{ \lambda }[/math] affects recBC enzyme in E. Coli. Karu 1975
[math]\displaystyle{ \gamma }[/math] protein of phage [math]\displaystyle{ \lambda }[/math] inhibits recombination activites of RecBCD enzyme. Murphy 1991