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Methods Mol Biol. 2018;1830:325-335. doi: 10.1007/978-1-4939-8657-6_19.

In Situ Proximity Ligation Assay to Detect the Interaction Between Plant Transcription Factors and Other Regulatory Proteins.

Xu Y1,2, Gan ES1, Ito T3,4,5.

Author information

1
Temasek Life Sciences Laboratory (TLL), 1 Research Link, National University of Singapore, Singapore, Singapore.
2
Biological Sciences, Nara Institute of Science and Technology, Ikoma, Nara, Japan.
3
Temasek Life Sciences Laboratory (TLL), 1 Research Link, National University of Singapore, Singapore, Singapore. itot@bs.naist.jp.
4
Department of Biological Sciences, Faculty of Science, National University of Singapore, Singapore, Singapore. itot@bs.naist.jp.
5
Biological Sciences, Nara Institute of Science and Technology, Ikoma, Nara, Japan. itot@bs.naist.jp.

Abstract

In plants, a lot of transcription factors fulfill their roles in gene regulation through the interaction with other regulatory proteins and co-factors. Thus, confirmation of protein-protein interaction is key to understand the precise function of transcription factors. Many methods have been developed to investigate the protein-protein interaction in vivo and in vitro. In situ Proximity Ligation Assay (PLA) is an innovative method to test protein-protein interaction in your tissues or cells of interest in vivo. Furthermore, this method allows us to detect transient interaction and low-abundance protein interaction with single molecule resolution. In this chapter, we describe a detailed protocol for the study of interaction between plant transcription factors and other regulatory proteins, in the scale of single nuclei of plant organ, tissues and cells.

KEYWORDS:

In situ proximity ligation assay; Nuclei; Protein-protein interaction; Tissues; Transcription factor

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