Quantification of RNA Content in Reconstituted Ebola Virus Nucleocapsids by Immunoprecipitation

Logan Banadyga; Hideki Ebihara

doi:10.1007/978-1-4939-7116-9_7

Quantification of RNA Content in Reconstituted Ebola Virus Nucleocapsids by Immunoprecipitation

Methods Mol Biol. 2017:1628:93-107. doi: 10.1007/978-1-4939-7116-9_7.

Authors

Logan Banadyga¹, Hideki Ebihara^{2

3}

Affiliations

¹ Laboratory of Virology, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT, USA.
² Laboratory of Virology, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT, USA. [email protected].
³ Department of Molecular Medicine, Mayo Clinic, Rochester, MN, USA. [email protected].

PMID: 28573613
DOI: 10.1007/978-1-4939-7116-9_7

Abstract

Immunoprecipitations are commonly used to isolate proteins or protein complexes and assess protein-protein interactions; however, they can also be used to assess protein-RNA complexes. Here we describe an adapted RNA immunoprecipitation technique that permits the quantification of RNA content in Ebola virus nucleocapsids that have been reconstituted in vitro by transient transfection.

Keywords: Ebola virus; Immunoprecipitation; Nucleocapsid; Nucleoprotein; RNA; VP24; VP35; qRT-PCR.

Publication types

Research Support, N.I.H., Intramural

MeSH terms

Ebolavirus / genetics*
Ebolavirus / pathogenicity
Hemorrhagic Fever, Ebola / genetics*
Hemorrhagic Fever, Ebola / virology
Humans
Immunoprecipitation / methods*
Nucleocapsid / chemistry
Nucleocapsid / genetics
Nucleocapsid / isolation & purification*
Nucleoproteins / genetics
Nucleoproteins / isolation & purification
RNA, Viral / genetics
RNA, Viral / isolation & purification
Virus Replication / genetics

Substances

Nucleoproteins
RNA, Viral