Agrobacterium transformation

Zhixiang Chen zhixiang at purdue.edu
Fri Jan 2 14:57:29 EST 2004


Dear Colleagues:

We are trying to use the series of binary vectors (pAOV, pKMB, pSOV etc.)
developed by Joshua Mylne in Australia (Plant Molecular Biology 1998,
16:257-262) for constitutive expression of Arabidopsis genes and are having
problem transforming them into the GV3101 strain of agrobacterium.  These
vectors were constructed using pSLJ75515 as backbone and are supposed to
have tet resistance in both E. coli and agrobacterium.  We have tried direct
transformation (using competent agrobacterial cells) and tri-parental mating
but couldn't get any positive transformant.  We would greatly appreciate it
if someone there who has used these vectors or the related pSLJ75515 could
share with us the detailed protocol of transforming these vectors into
GV3101 (including the medium and antibiotic used,their concentrations and
culture conditions, length etc.).  Thank you very much for your help.


Zhixiang Chen, Associate Professor
Department of Botany & Plant Pathology
915 West State Street
Purdue University
West Lafayette, IN 47907-2054

Phone: 765-494-4657
FAX:  765-494-5896
E-mail: zhixiang at purdue.edu


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