nos promoter in Agro

jkreps at biochem.umass.edu jkreps at biochem.umass.edu
Thu Oct 26 15:26:06 EST 1995


>Return-path: <BIOSCI-REQUEST at net.bio.net>
>Date: Thu, 26 Oct 1995 17:45:36 +0000
>From: Yves.Poirier at ibpv.unil.ch
>Subject: nos promoter in Agro
>Sender: lpddist at mserv1.dl.ac.uk
>To: arabidopsis at net.bio.net
>Original-To: arab-gen at dl.ac.uk
>
>Dear netters:
>
>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?
>Thank you
>Yves Poirier
>Institut of Plant Biology
>University of Lausanne
>
>
>
>
Dear Yves and other Netters,

Informally, I can say that as a grad. student working with a cosmid that had
the npt II gene driven by nos and a carbenicillin-R gene for bacterial
selection, I could get single colonies growing on Kan, either in E.coli. or
in Agrobacterium.  The growth I saw was less than a normal KanR bacterium
when compared side by side, however, a control strain lacking the cosmid did
not grow at all.

Joel A. Kreps
Dept. of Biochem and Mol Bio
UMass at Amherst
Massachusetts, USA




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