Abstract
The study of protein–protein interactions is critical for the understanding and regulation of biological systems. To that end, yeast two-hybrid systems have been used to study protein–protein interactions in vivo, but they frequently suffer from a high incidence of false positives when applied to mammalian systems. A novel mammalian two-hybrid system has recently been developed which exhibits lower background and higher sensitivity than earlier mammalian two-hybrid systems. It has successfully detected interactions with dissociation constants ranging from 0.99 nM to 55 μM. The system was built upon the tetracycline repressor–tetracycline operator interaction and is suitable for use in the study of most, if not all, mammalian protein–protein interactions.
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Acknowledgments
The authors would like to thank Gabrielle Thibodeaux and Roshani Cowmeadow for initial development of the trM2H system, and Aiko Umeda, Jie Zhu, Liang Xiang, YungAh Lee, Wonjae Lee, Jay Xiao, Kristi Kim, Young Park, and Sarah Chun for technical assistance.
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© 2012 Springer Science+Business Media, LLC
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Moncivais, K., Zhang, Z.J. (2012). Tetracycline Repressor-Based Mammalian Two-Hybrid Systems. In: Suter, B., Wanker, E. (eds) Two Hybrid Technologies. Methods in Molecular Biology, vol 812. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-455-1_15
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DOI: https://doi.org/10.1007/978-1-61779-455-1_15
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Publisher Name: Humana Press, Totowa, NJ
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