Basta-only plant vector ?
bjanssen at bio.tamu.edu
Mon Dec 4 17:53:01 EST 1995
In article <49uesh$bp9 at mserv1.dl.ac.uk>, vachon at bio.grenet.fr (Gilles
> Hi all,
> Some of you might recall my message about 35S-Nos' vectors some months ago.
> I received several answers and vectors too...(Thanks to Reinhard Hehl and
> David Oppenheimer and many others....).
> I now have another request about plant vectors.
> For genetic reasons, we have to make transgenic lines with anything but
> Kanamycine resistance (or no resistance cassette at all...).
> The best for us would be a Basta-only resistance vector.
> Has someone have to deal with the same problem and made such a vector already?
I have a set of binary vectors with the general structure:
LB ocs3'PacI selectable marker AscI nos5' or 35S5' lacz RB
then using a shuttle vector I have cloned in the bar gene, the nptII gene
and the hpt gene. So I have vectors that are essentially identical but
have different selectable marker genes (I have all three 35S versions but
only nptII and hpt driven by the nos5').
I have also made versions of these vectors that have a 35S 5' intron GUS
ocs3' cloned into the unique NotI site in the lacz, these have been used
to develop transformation systems. A number of shuttle vectors that clone
into the unique NotI site in the lacz have also been made including:
NotI-MCS-NotI; NotI-35S5'-MCS-ocs3'-NotI; and NotI-MCS-GUS-ocs3'-NotI
(with all three reading frames so that tracriptional and translational
promoter GUS fusions can be made). Because the lacz has a functional
blue/white screen cloning into the binary is relatively easy and cloning
into the "small" shuttle vectors is also relatively easy.
All of these vectors and shuttles are freely available.
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