nos promoter in Agro

Yasuo Niwa niwa at FNS1.U-SHIZUOKA-KEN.AC.JP
Sun Oct 29 22:18:21 EST 1995


At  5:45 PM 95.10.26 +0000, <Yves.Poirier at ibpv.unil.ch> wrote:
>
>Does anybody knows if the nos promoter is active in Agrobacterium. We are
>using a binary vector which has a nptII gene driven by the nos promoter for
>selection of plants on kanamycin. The vector has strep/spec resistance for
>selection of Agrobacterium and E. coli containing the plasmid. In one
>experiment, we transfected the Agro (which is kanamycin sensitive but
>gentamycin resistant) with the vector and selected by mistake with
>kanamycin and gentamycin...but the Agrobacterium grew well. We later
>checked and the right plasmid was present in Agro. The only explanation I
>have right now is that the nos promoter is active in Agro. Is this a known
>fact? If so, could somebody point a refernce to me?

E. coli and Agrobacterium carring the pGA482 binary vector (nos-nptII) can 
be maintained with tet and 10 micro g/ml of kanamycin in their culture 
medium.               Methods in Enzymology vol.153 p294

The concentration of kan depends on the type of nos promoter and ori.
***************************************
       Yasuo Niwa, Ph. D.
 Laboratory of Plant Cell Technology
   University of Shizuoka
 52-1 Yada, Shizuoka 422, Japan
 Phone&Fax: +81(Japan)-54-264-5582
 e-mail: niwa at fns1.u-shizuoka-ken.ac.jp
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