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BN-PAGE (Blue Native-Polyacrylamide Gel Electrophoresis) is a non-denaturing electrophoretic technique used to analyze the molecular weight and oligomeric states of protein complexes under near-native conditions. NLRs (nucleotide-binding leucine-rich repeat proteins), which function as intracellular immune receptors in plants, form oligomeric higher-order complexes known as resistosomes upon activation by recognition of pathogen effectors—a mechanism elucidated through BN-PAGE and structural analyses. Here, we describe a method combining BN-PAGE with Agrobacterium-mediated complementation assay to investigate the resistosome formation of the NLR protein ZAR1 in Nicotiana benthamiana.

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Blue Native-PAGE Assay of NLR Protein Complexes Combined with Genetic Complementation in Nicotiana benthamiana

  • 实验方案

  • 发表日期：

    2026年

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  • Kodai Honda,
  • Tadashi Fujiwara,
  • Hiroaki Adachi

摘要

BN-PAGE (Blue Native-Polyacrylamide Gel Electrophoresis) is a non-denaturing electrophoretic technique used to analyze the molecular weight and oligomeric states of protein complexes under near-native conditions. NLRs (nucleotide-binding leucine-rich repeat proteins), which function as intracellular immune receptors in plants, form oligomeric higher-order complexes known as resistosomes upon activation by recognition of pathogen effectors—a mechanism elucidated through BN-PAGE and structural analyses. Here, we describe a method combining BN-PAGE with Agrobacterium-mediated complementation assay to investigate the resistosome formation of the NLR protein ZAR1 in Nicotiana benthamiana.