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Two-hybrid assay in plants

In the August issue of Nature Biotechnology, Rajagopal Subramaniam and colleagues from the Université de Montreal describe a system for visualizing protein-protein interactions in living plant cells (Nature Biotechnology 2001, 19:769-772). Their technique is based on an in vivoprotein-fragment complementation assay in which protein interactions between fusion proteins induce folding and reassembly of fragments of the murine dihydrofolate reductase (DHFR) enzyme. Protein-protein interactions are monitored by the DHFR inhibitor fluorescein-conjugated methotrexate (fMTX). Subramaniam et al. used their assay to examine the interaction between Arabidopsis disease-resistance protein NPR1/NIM1 and the basic leucine-zipper protein TGA2. They show that salicylic acid induced NPR1-TGA2 interaction in tobacco or potato cells. This plant two-hybrid system should prove useful for the functional annotation of plant genomes.


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    Nature Biotechnology, []

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    Université de Montreal , []

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    Clonal selection and in vivo quantitation of protein interactions with protein-fragment complementation assays.

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    The Arabidopsis NPR1 gene that controls systemic acquired resistance encodes a novel protein containing ankyrin repeats

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Weitzman, J.B. Two-hybrid assay in plants. Genome Biol 2, spotlight-20010807-01 (2001).

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  • Methotrexate
  • Fusion Protein
  • Salicylic Acid
  • Functional Annotation
  • Plant Genome