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.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Phizicky, E.M. and S. Fields (1995) Protein-protein interactions: methods for detection and analysis Microbiol Rev. 59, 94–123
Luo, Y., et al. (1997) Mammalian two-hybrid system: A complementary approach to the yeast two-hybrid system Biotechniques 22, 350–352
Thibodeaux, G.N., et al. (2009) A tetracycline repressor-based mammalian two-hybrid system to detect protein-protein interactions in vivo Anal Biochem. 386, 129–31
Huang H., J.B.M., Bader J.S. (2007) Where have all the interactions gone? Estimating the coverage of two-hybrid protein interac-tion maps PLoS Computational Biology 3, 2155–2174
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.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
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
Download citation
DOI: https://doi.org/10.1007/978-1-61779-455-1_15
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-454-4
Online ISBN: 978-1-61779-455-1
eBook Packages: Springer Protocols