Abstract
Yeast two-hybrid systems are powerful tools to identify novel protein–protein interactions and have been extensively used to study viral protein interactions. The most commonly used systems are GAL4-based and LexA-based systems. Over the last decade, a range of modifications and improvements have been made to the original yeast two-hybrid system to expand the scope of molecular interaction assays and to eliminate false positives. Detailed protocols are provided for yeast strain storage, yeast transformation, yeast mating, preparation of growth and selection medium, quantitative reporter gene assays (α- and β-galactosidase liquid assays) and detection of fusion protein by Western blot.
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Acknowledgments
We thank Dr Qingzhen Liu for critically reading the manuscript and Xiaoyun Wu for compiling the reference list. Deyin Guo's laboratory is supported by the Luojia Professorship Program of Wuhan University and China National Science Foundation (NSFC-30570394 and 30270313). The YTHS studies on plant virus proteins of M.L. Rajamäki and J.P.T. Valkonen are funded by the Academy of Finland (grant 204104, 1102003, 1102134 and 1110797).
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Guo, D., Rajamäki, ML., Valkonen, J. (2008). Protein–Protein Interactions: The Yeast Two-Hybrid System. In: Foster, G.D., Johansen, I.E., Hong, Y., Nagy, P.D. (eds) Plant Virology Protocols. Methods in Molecular Biology™, vol 451. Humana Press. https://doi.org/10.1007/978-1-59745-102-4_29
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DOI: https://doi.org/10.1007/978-1-59745-102-4_29
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