利用改良的Gateway兼容载体进行双分子荧光互补以可视化植物细胞中的蛋白质-蛋白质相互作用

Bimolecular Fluorescence Complementation with Improved Gateway-Compatible Vectors to Visualize Protein-Protein Interactions in Plant Cells.

作者信息

Goto-Yamada Shino, Hikino Kazumi, Nishimura Mikio, Nakagawa Tsuyoshi, Mano Shoji

机构信息

Malopolska Centre of Biotechnology, Jagiellonian University, Krakow, Poland.

Department of Evolutionary Biology and Biodiversity, National Institute for Basic Biology, Okazaki, Japan.

出版信息

Methods Mol Biol. 2018;1794:245-258. doi: 10.1007/978-1-4939-7871-7_16.

DOI:10.1007/978-1-4939-7871-7_16

PMID:29855962

Abstract

The bimolecular fluorescence complementation (BiFC) assay is a powerful, flexible, and simple tool to study protein-protein interactions in living cells. To accelerate the production and assessment of BiFC constructs, Gateway-compatible multicolor BiFC vectors were generated to enable the simultaneous production of multiple fusion genes that have the split N- or C-terminal fragment of fluorescent protein with the gene of interest in a high-throughput manner. Two different transient expression techniques for the assessment of BiFC in plant cells are described.

摘要

双分子荧光互补（BiFC）分析是一种用于研究活细胞中蛋白质-蛋白质相互作用的强大、灵活且简单的工具。为了加速BiFC构建体的产生和评估，构建了与Gateway兼容的多色BiFC载体，以高通量方式同时产生多个融合基因，这些融合基因将荧光蛋白的N端或C端片段与感兴趣的基因分开。本文描述了两种用于评估植物细胞中BiFC的不同瞬时表达技术。