Abstract
Yeast two-hybrid (Y2H) is a technique used to identify protein-protein interactions. It relies on interacting proteins bringing the two domains of a split transcription factor into close proximity, thereby reconstituting its ability to activate reporter genes. There are many variations on this technique. Here we provide an adapted protocol based on the Invitrogen ProQuest™ Two-Hybrid system, which has been used successfully to screen over 60 Phytophthora infestans pathogen effector proteins to identify candidate-interacting proteins in Solanum tuberosum, and has been used to identify many potato-potato protein-protein interactions.
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References
Invitrogen ProQuest™ 2-Hybrid System: a sensitive method for detecting protein-protein interactions (2005) User manual
Vidal M (1997) The reverse two-hybrid system. In: The yeast two-hybrid system. Oxford University Press, Oxford, pp 109–147
Boevink P, McLellan H, Bukharova T, Engelhardt S, Birch P (2014) In vivo protein–protein interaction studies with BiFC: conditions, cautions, and caveats. In: Plant-pathogen interactions. Humana Press, Totowa, NJ, pp 81–90
Acknowledgments
This work was supported by BBSRC grants BB/P020569/1, BB/N009967/1, BB/K018183/1, and BB/G015244/1 to P.R.J.B.
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McLellan, H., Armstrong, M.R., Birch, P.R.J. (2021). Yeast Two-Hybrid Screening for Identification of Protein-Protein Interactions in Solanum tuberosum . In: Dobnik, D., Gruden, K., Ramšak, Ž., Coll, A. (eds) Solanum tuberosum. Methods in Molecular Biology, vol 2354. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1609-3_4
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DOI: https://doi.org/10.1007/978-1-0716-1609-3_4
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1608-6
Online ISBN: 978-1-0716-1609-3
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