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Selection of Arabidopsis mutants overexpressing genes driven by the promoter of an auxin-inducible glutathione S-transferase gene

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Author: Kop, D.A.M. van der · Schuyer, M. · Pinas, J.E. · Zaal, B.J. van der · Hooykaas, P.J.J.
Institution: Centraal Instituut voor Voedingsonderzoek TNO
Source:Plant Molecular Biology, 5, 39, 979-990
Identifier: 86824
doi: doi:10.1023/A:1006129426712
Keywords: Nutrition · Arabidopsis thaliana · Auxin-responsive gene expression · Developmental mutants · Glutathione S-transferase · GST · Gup mutant · Auxin · Beta glucuronidase · Gene expression · Glutathione transferase · Kanamycin · Reporter gene · Transgenic plant · Arabidopsis · Chromosome Mapping · Crosses, Genetic · DNA, Bacterial · Enzyme Induction · Ethyl Methanesulfonate · Gene Expression Regulation, Plant · Genes, Plant · Glutathione Transferase · Indoleacetic Acids · Mutagenesis · Mutagens · Mutation · Phenotype · Plants, Genetically Modified · Promoter Regions (Genetics) · Recombinant Fusion Proteins · Up-Regulation


Transgenic arabidopsis plants were isolated that contained a T-DNA construct in which the promoter of an auxin-inducible glutathione S-transferase (GST) gene from tobacco was fused to the kanamycin resistance (nptII) as well as to the β-glucuronidase (gusA) reporter gene. Subsequently, seeds were treated with EMS to obtain mutants in which both reporter gene fusions were up-regulated. Northern analysis showed that the mRNA level of a related, endogenous auxin-inducible GST gene of Arabidopsis was increased in some of these mutants as well. Two of the gup (GST up-regulated) mutants were characterized in more detail and roughly mapped. Both had epinastic cotyledons and leaves, a phenotype that turned out to be linked to the gup mutation. Chemicals/CAS: DNA, Bacterial; Ethyl Methanesulfonate, 62-50-0; Glutathione Transferase, EC; Indoleacetic Acids; Mutagens; Recombinant Fusion Proteins; T-DNA