A field-proven yeast two-hybrid protocol used to identify coronavirus-host protein-protein interactions
Vidalain, Pierre Olivier; Jacob, Yves; Hagemeijer, Marne C.; Jones, Louis M.; Neveu, Grégory; Roussarie, Jean Pierre; Rottier, Peter J M; Tangy, Frédéric; De Haan, Cornelis A M
(2015) Coronaviruses: Methods and Protocols, pp. 213 - 229Methods in Molecular Biology, pp. 213 - 229
(Part of book)
Abstract
Over the last 2 decades, yeast two-hybrid became an invaluable technique to decipher protein-protein interaction networks. In the field of virology, it has proven instrumental to identify virus-host interactions that are involved in viral embezzlement of cellular functions and inhibition of immune mechanisms. Here, we present a yeast two-hybrid protocol
... read more
that has been used in our laboratory since 2006 to search for cellular partners of more than 300 viral proteins. Our aim was to develop a robust and straightforward pipeline, which minimizes false-positive interactions with a decent coverage of target cDNA libraries, and only requires a minimum of equipment. We also discuss reasons that motivated our technical choices and compromises that had to be made. This protocol has been used to screen most non-structural proteins of murine hepatitis virus (MHV), a member of betacoronavirus genus, against a mouse brain cDNA library. Typical results were obtained and are presented in this report.
show less
Download/Full Text
Not available. The author may have various reasons for not providing access, for instance because it is prohibited by the commissioner of the research, or because the author is conducting further research on the subject.
Keywords: Host-pathogen interactions, Interactomics, Murine hepatitis virus, Proteomics, Yeast two-hybrid, Immunology and Microbiology(all), Medicine(all), Journal Article, Research Support, Non-U.S. Gov't
ISSN: 1064-3745
ISBN: 9781493924370
9781493924387
Publisher: Springer New York
(Publisher version, Peer reviewed)
See more statistics about this item