@inbook{ba32d4f56b69436a89e52281e45f12ab,
title = "Preparation of recombinant alphaviruses for functional studies of ADP-ribosylation",
abstract = " Recently we characterized the mono(ADP-ribosyl) hydrolase (MAR hydrolase) activity of the macrodomain of nonstructural protein 3 (nsP3 MD ) of chikungunya virus. Using recombinant viruses with targeted mutations in the macrodomain, we demonstrated that hydrolase function is important for viral replication in cultured neuronal cells and for neurovirulence in mice. Here, we describe the general cell culture and animal model infection protocols for alphaviruses and the technical details for biochemical characterization of the MAR hydrolase activity of nsP3 MD mutants and the preparation of recombinant viruses incorporating those mutations through site-directed mutagenesis of an infectious cDNA virus clone.",
keywords = "Alphavirus, Chikungunya virus, In vitro transcription, Infectious cDNA clone, MAR hydrolase assay, Reverse genetics, Sindbis virus, Site-directed mutagenesis",
author = "Rachy Abraham and McPherson, {Robert Lyle} and Easwaran Sreekumar and Leung, {Anthony K.L.} and Griffin, {Diane E.}",
note = "Publisher Copyright: {\textcopyright} 2018, Springer Science+Business Media, LLC, part of Springer Nature.",
year = "2018",
doi = "10.1007/978-1-4939-8588-3_21",
language = "English (US)",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "297--316",
booktitle = "Methods in Molecular Biology",
}