Curing E.coli plasmids

Rafael Maldonado rafael at corona.med.utah.edu
Mon Mar 20 14:33:46 EST 1995


On Mon, 20 Mar 1995, Duncan Clark wrote:

> Hi Folks,
> 
> Has anyone got a good high efficiency method for curing both colE1 
> replicon based plasmids (pBR322 deriv.) and p15A replicon based 
> plasmids (pACYC series). EtBr, Acridine Orange, electroporation 
> (if plasmids can go in they can also come out!) etc.

You can use coumermycin. It's a inhibitor of the DNA gyrase and the 
pasmids are lost at sub-lethal concentrations. I used 5 
ug/ml and it works well with pBR322-derivated plasmid. You can use it for 
others replicons as well.

The refrence is:
Danielvskaya, ON and Gragerov, AI (1980). Curing of Escherichia coli K12 
plasmids by coumermycin. MGG 178, 233-235.

You can find coumermycin at Sigma (C-9270). Not relation with the company!


Rafa



__________________________________________________________________________

Rafael Maldonado			"You gave to a boy the job  
Department of Human Genetics		 of a man. He's dead."
University of Utah			
Salt Lake City, Utah 84112. USA.	                     John Wayne
Rafael at genetics.med.utah.edu
Rafael at corona.med.utah.edu		"No por mucho madrugar, amanece
Tlf: 801-581-4429			mas temprano"
Fax: 801-585-3910			
					                  Saber popular





More information about the Methods mailing list