Protein-Protein Interaction via Two-Hybrid Assay in Yeast.

Christiane Eliza Motta Duarte, Nivea Costa Euclydes

Methods in molecular biology (Clifton, N.J.) 2024

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The yeast two-hybrid assay enables detecting interactions between proteins, which makes this tool of particular interest for plant-virus interaction studies. Basically, the reporter gene expression (HIS3) is activated by the binding of a transcription factor GAL4, which, like eukaryotic transcription factors, is modular in nature and consists of two structurally independent domains: DNA-binding (DB) and activation (AD) domains. The two proteins under investigation are expressed separately, one fused to the BD domain and the other to the AD domain. In the yeast strain AH109, activation of the reporter gene occurs only in cells that contain interacting proteins, reconstituting the transcription factor GAL4 which then binds to the responsive promoter and results in yeast colony growth. © 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Citation

Christiane Eliza Motta Duarte, Nivea Costa Euclydes. Protein-Protein Interaction via Two-Hybrid Assay in Yeast. Methods in molecular biology (Clifton, N.J.). 2024;2724:193-210