We have developed a simple and totally in vitro selection procedure based on cell-free cotranslation using a highly stable and efficient in vitro virus (IVV). Cell-free cotranslation of tagged bait and prey proteins is advantageous for the formation of protein complexes and allows high-throughput analysis of protein–protein interactions (PPI) as a result of providing in vitro instead of in vivo preparation of bait proteins. The use of plural selection rounds and a two-step purification of the IVV selection, followed by in vitro post-selection, is advantageous for decreasing false positives. This simple IVV selection system based on cell-free cotranslation is applicable to high-throughput and comprehensive analysis of transcription factor networks.
CITATION STYLE
Ohashi, H., Fujimori, S., Hirai, N., Yanagawa, H., & Miyamoto-Sato, E. (2014). Analysis of transcription factor networks using IVV method. Methods in Molecular Biology, 1164, 15–22. https://doi.org/10.1007/978-1-4939-0805-9_2
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