植物转录因子与DNA互作研究技术

doi:10.11983/CBB20057

植物学报 ›› 2020, Vol. 55 ›› Issue (4): 468-474.DOI: 10.11983/CBB20057 cstr: 32102.14.CBB20057

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()

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()

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/Abstract

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

杨立文,刘双荣,李玉红,林荣呈. 植物转录因子与DNA互作研究技术. 植物学报, 2020, 55(4): 468-474.

Liwen Yang,Shuangrong Liu,Yuhong Li,Rongcheng Lin. Methods for Examining Transcription Factor-DNA Interaction in Plants. Chinese Bulletin of Botany, 2020, 55(4): 468-474.

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链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB20057

https://www.chinbullbotany.com/CN/Y2020/V55/I4/468

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参考文献 16

[1]	杨立文, 刘双荣, 林荣呈 (2019). 光信号与激素调控种子休眠和萌发研究进展. 植物学报 54, 569-581.
[2]	Chen DQ, Xu G, Tang WJ, Jing YJ, Ji Q, Fei ZJ, Lin RC (2013). Antagonistic Basic Helix-Loop-Helix/bZIP transcription factors form transcriptional modules that integrate light and reactive oxygen species signaling in Arabidopsis. Plant Cell 25, 1657-1673. DOI URL PMID
[3]	Cheng MC, Enderle B, Kathare PK, Islam R, Hiltbrunner A, Huq E (2020). PCH1 and PCHL directly interact with PIF1, promote its degradation, and inhibit its transcriptional function during photomorphogenesis. Mol Plant 13, 499-514. DOI URL PMID
[4]	Hellens RP, Allan AC, Friel EN, Bolitho K, Grafton K, Templeton MD, Karunauretnam S, Gleave AP, Laing WA (2005). Transient expression vectors for functional genomics, quantification of promoter activity and RNA silencing in plants. Plant Methods 1, 13. URL PMID
[5]	Jiang ZM, Xu G, Jing YJ, Tang WJ, Lin RC (2016). Phytochrome B and REVEILLE1/2-mediated signaling controls seed dormancy and germination in Arabidopsis. Nat Commun 7, 12377. DOI URL PMID
[6]	Jing YJ, Guo Q, Lin RC (2019). The chromatin-remodeling factor PICKLE antagonizes polycomb repression of FT to promote flowering. Plant Physiol 181, 656-668. DOI URL PMID
[7]	Latchman DS (2005). Gene Regulation-A Eukaryotic Perspective, 5th edn. Oxford and New York: Taylor and Fran-cis. pp. 1.
[8]	Li JJ, Herskowitz I (1993). Isolation of ORC6, a component of the yeast origin recognition complex by a one-hybrid system. Science 262, 1870-1874. URL PMID
[9]	Lin RC, Ding L, Casola C, Ripoll DR, Feschotte C, Wang HY (2007). Transposase-derived transcription factors regulate light signaling in Arabidopsis. Science 318, 1302-1305. URL PMID
[10]	Nakashima K, Yamaguchi-Shinozaki K, Shinozaki K (2014). The transcriptional regulatory network in the drought response and its crosstalk in abiotic stress responses including drought, cold, and heat. Front Plant Sci 5, 170. DOI URL PMID
[11]	Ohama N, Sato H, Shinozaki K, Yamaguchi-Shinozaki K (2017). Transcriptional regulatory network of plant heat stress response. Trends Plant Sci 22, 53-65. URL PMID
[12]	Pu L, Brady S (2010). Systems biology update: cell type- specific transcriptional regulatory networks. Plant Physiol 152, 411-419. DOI URL PMID
[13]	Wu FH, Shen SC, Lee LY, Lee SH, Chan MT, Lin CS (2009). Tape-Arabidopsis Sandwich-a simpler Arabidopsis protoplast isolation method. Plant Methods 5, 16. URL PMID
[14]	Xu G, Jiang ZM, Wang HY, Lin RC (2019). The central circadian clock proteins CCA1 and LHY regulate iron homeostasis in Arabidopsis. J Integr Plant Biol 61, 168-181. URL PMID
[15]	Yang LW, Jiang ZM, Liu SR, Lin RC (2020). REVEILLE1 inhibits RGL2 degradation to regulate seed dormancy and germination in Arabidopsis. New Phytol 225, 1593-1605. URL PMID
[16]	Yoo SD, Cho YH, Sheen J (2007). Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis. Nat Protoc 2, 1565-1572. DOI URL PMID

植物转录因子与DNA互作研究技术

Methods for Examining Transcription Factor-DNA Interaction in Plants

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