Protocol to identify protein-protein interaction networks in Solanum tuberosum using transient TurboID-based proximity labeling

使用基于 TurboID 的瞬时邻近标记法识别马铃薯中的蛋白质 - 蛋白质相互作用网络的协议

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作者:Li Shi, Tatiana Marti Ferrando, Sergio Landeo Villanueva, Matthieu H A J Joosten, Vivianne G A A Vleeshouwers, Christian W B Bachem

Abstract

Protein-protein interactions (PPIs) in crop plants remain largely unexplored. Here, we provide a protocol for identifying PPIs in potato (Solanum tuberosum) using TurboID-mediated proximity labeling. We transiently expressed constructs for a nucleus-located transcription factor and a plasma membrane-localized receptor-like kinase fused to TurboID to identify PPIs in potato leaves. We describe the plasmid construction, plant material, agroinfiltration, biotin treatment, protein isolation, free biotin removal, western blot analysis, and enrichment of biotinylated proteins for mass spectrometry analysis.

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