Basta-only plant vector ?

Bart Janssen 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
VACHON) wrote:

> 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?

Yes
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.

cheers
Bart



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