植物学报 ›› 2020, Vol. 55 ›› Issue (4): 468-474.DOI: 10.11983/CBB20057

• 特邀专家方法 • 上一篇    下一篇

植物转录因子与DNA互作研究技术

杨立文,刘双荣,李玉红,林荣呈()   

  1. 中国科学院植物研究所光生物学重点实验室, 北京 100093
  • 收稿日期:2020-04-04 接受日期:2020-06-28 出版日期:2020-07-01 发布日期:2020-06-29
  • 通讯作者: 林荣呈
  • 基金资助:
    国家自然科学基金(31870213)

Methods for Examining Transcription Factor-DNA Interaction in Plants

Liwen Yang,Shuangrong Liu,Yuhong Li,Rongcheng Lin()   

  1. Key Laboratory of Photobiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China
  • Received:2020-04-04 Accepted:2020-06-28 Online:2020-07-01 Published:2020-06-29
  • Contact: Rongcheng Lin

摘要: 转录通过调控下游基因的时空特异性表达影响植物生长发育。在转录调控机制的解析过程中, 转录因子与DNA的相互作用是关键的一环。近年来, 研究者利用酵母单杂交(Y1H)和凝胶阻滞迁移率实验(EMSA)检测转录因子能否直接结合DNA; 而瞬时表达技术则是一种检测转录因子对下游基因调控作用的便捷方式。该文对Y1H、EMSA和瞬时表达技术的原理、实验方法和相关注意事项进行详细阐述, 以期为转录因子与DNA的互作研究提供参考方法。

关键词: 转录因子, DNA, Y1H, EMSA, 瞬时表达技术

Abstract: Transcription affects the growth and development of plants through regulating the spatio-temporal expression of downstream genes. The interaction between transcription factors and DNA is a key section in the process of exploring transcriptional regulatory networks. In the past few years, researchers utilize yeast one hybrid (Y1H) and electrophoresis mobility shift assay (EMSA) to examine whether a transcription factor directly interacts with target DNA. In addition, transient luciferase activity assay provides a convenient method for researchers to test the regulation of transcription factors on downstream gene expression. In this paper, we elaborate the principles, methods, and advantages and limitations of Y1H, EMSA and transient luciferase activity assay, to provide technical references for exploring the transcription factor-DNA interactions.

Key words: transcription factor, DNA, Y1H, EMSA, transient luciferase activity assay