linear DNA Xformation

Tibor WEIS,network manager tibor at tuzvonet.so
Tue May 4 11:02:14 EST 1993


In article <C5r7yA.6vs at ncifcrf.gov> pnh at fcsparc6.ncifcrf.gov (Paul N Hengen) writes:
>Relay-Version: VMS News - V6.1B5 17/9/92 VAX/VMS V5.4-1; site mvax.uakom.cs
>Path: mvax.uakom.cs!aci.cvut.cs!alijku11!news.univie.ac.at!scsing.switch.ch!ira.uka.de!
> sol.ctr.columbia.edu!zaphod.mps.ohio-state.edu!darwin.sura.net!fconvx.ncifcrf.gov!fcsparc6!pnh
>Newsgroups: bionet.molbio.methds-reagnts
>Subject: Re: linear DNA Xformation
>Message-ID: <C5r7yA.6vs at ncifcrf.gov>
>From: pnh at fcsparc6.ncifcrf.gov (Paul N Hengen)
>Date: Mon, 19 Apr 1993 23:30:10 GMT
>Sender: Paul N. Hengen
>References: <1993Apr19.094503.26490 at gserv1.dl.ac.uk>
>Distribution: bionet
>Organization: Frederick Cancer Research and Development Center
>Summary: Reference
>Nntp-Posting-Host: fcsparc6.ncifcrf.gov
>Lines: 40
>
>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



More information about the Methods mailing list