Simple question that is hard to answer

peter via methods%40net.bio.net (by peter.ianakiev from gmail.com)
Fri Feb 16 09:38:41 EST 2007


On Feb 16, 7:38 am, ChenHA <h... from freeuk.com> wrote:
> On Thu, 15 Feb 2007 16:51:49 -0500, "Carlo Zambonelli"
>
> <zambo... from gmail.com> wrote:
> >Yes, as long as you have 2 different selective markers and the appropriate
> >cell strain. Seehttp://www.emdbiosciences.com/html/NVG/DuetTable.html
> >C.
>
> I think you misunderstood.  The Duet series are plasmids with
> different replicons, so are different as far as the original question
> is concerned.  The question would be one about plasmids with the same
> replicon, and that is problematic.  If there are different antibiotics
> used, then it may be possible to maintain the plasmids, but do
> remember that antibiotics like ampicillin are broken down fairly
> quickly, and when the antibiotics are gone, there is in fact no
> selection, and one of the plasmids will most probably be lost.
>
> I would suggest trying different plasmids for the experiment.  The
> Duet series would be a useful one since they are of different
> replicons, although they are designed for expressing multiple genes (2
> in each plasmids).   It is therefore possible to clone both genes into
> the same plasmids, although I'm wary of having 2 copies of essentially
> the same gene in one plasmids (possible deletion).  It might be
> cheaper and easier just to find someone who has a different plasmid,
> e.g. a pACYC based plasmids
>
>
>
> >On 15 Feb 2007 12:20:34 -0800, peter <peter.ianak... from gmail.com> wrote:
>
> >> Hi group,
> >> I have a question regarding incompatibility of plasmids. Can one
> >> bacterial cell be transformed with more than one plasmid (of the same
> >> kind) and keep these plasmids. For example if one has two clones (same
> >> insert, same vector) can they co-exist in the same cell (same size
> >> too, only difference would be point mutation). Your thoughts are
> >> invaluable and much appreciated.
> >> Peter
>
> >> _______________________________________________
> >> Methods mailing list
> >> Meth... from net.bio.net
> >>http://www.bio.net/biomail/listinfo/methods- Hide quoted text -
>
> - Show quoted text -

Thanks Chen,
I am not trying to clone anything. I am doing development work, in
which I wonder if I have mixed population of different point mutant
plasmids in same bacteria or one plasmid with different point
mutations. Simple question - hard to answer without doing experiment.
I thought someone might have the answer, but...



More information about the Methods mailing list