植物学报 ›› 2020, Vol. 55 ›› Issue (1): 69-75.DOI: 10.11983/CBB19229

• 技术方法 • 上一篇    下一篇

萤火素酶互补实验检测蛋白互作

赵燕(),周俭民   

  1. 中国科学院遗传与发育生物学研究所, 北京 100101
  • 收稿日期:2019-11-24 接受日期:2019-12-02 出版日期:2020-01-01 发布日期:2019-12-20
  • 通讯作者: 赵燕
  • 基金资助:
    国家自然科学基金(31471741)

Luciferase Complementation Assay for Detecting Protein Interactions

Yan Zhao(),Jianmin Zhou   

  1. Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China
  • Received:2019-11-24 Accepted:2019-12-02 Online:2020-01-01 Published:2019-12-20
  • Contact: Yan Zhao

摘要:

蛋白质-蛋白质相互作用在真核生物的各项生命活动中发挥重要作用。与其它蛋白质互作研究技术相比, 借助于烟草(Nicotiana benthamiana)瞬时表达系统的萤火素酶互补实验(LCA)具有简单、灵敏、可靠、高效和低背景等优点, 并可轻松扩展为大规模蛋白质互作的筛选和验证研究。该文介绍了萤火素酶互补实验的具体操作过程, 通过2种数据收集方法来定性并定量分析生物发光或发光强度, 从而检测植物目标蛋白之间的相互作用。

关键词: 萤火素酶互补实验, 蛋白质相互作用, 烟草瞬时表达

Abstract:

Protein-Protein interactions play important roles in various eukaryotic biological processes. Compared to other techniques measuring protein-protein interactions in plants, the Luciferase Complementation Assay (LCA), based on Agrobacterium-mediated transient expression in Nicotiana benthamiana, is a simple, sensitive, reliable, highly quantitative and low background method that can be easily scaled up for high-throughput interactome studies. Here, we describe a protocol that includes two alternative data collection methods to qualitative and quantitative analyse luminescence or luminous intensity to detect protein-protein interactions in plant cells.

Key words: luciferase complementation assay, protein-protein interactions, tobacco transient expression