linear DNA Xformation

Paul N Hengen pnh at fcsparc6.ncifcrf.gov
Mon Apr 19 18:30:10 EST 1993


IBELGAUFTS at de.mpg.biochem.vms wrote:

: Are there any reasons why transformation of bacteria with linear DNA
: should not work? Does it work?

Mike Poidinger wrote:

> It depends on why you want to do it.
 
> My understanding is that Eukaryotic Xformations are done 
> with linear DNA to improve the eficiency of integration into 
> the chromosome. Since most bacterial Xformations involve
> autonomously replicating plasmids, linearizing the 
> DNA would not serve much purpose. And if you actually wanted
> chromosomal integration, I would guess there are better ways
> than Xformation with linear DNA and then hoping :-)

It could be you would like to force an integration or perhaps
cause a specific mutation within the bacterial chromosome.

See the following article for a method to do such a thing:

@article{Winans1985, 
author = "S. C. Winans
     and S. J. Elledge
     and J. H. Krueger
     and G. C. Walker",
title = "Site-directed insertion and deletion mutagenesis
with cloned fragments in Escherichia coli",
journal = "J. Bacteriol.",
volume = "161",
pages = "1219-1221",
year = "1985"}


Paul N. Hengen
National Cancer Institute
Frederick Cancer Research and Development Center
Frederick, Maryland 21702-1201 USA



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