Modeling Ebola Virus Genome Replication and Transcription with Minigenome Systems

Tessa Cressey; Kristina Brauburger; Elke Mühlberger

doi:10.1007/978-1-4939-7116-9_6

Modeling Ebola Virus Genome Replication and Transcription with Minigenome Systems

Methods Mol Biol. 2017:1628:79-92. doi: 10.1007/978-1-4939-7116-9_6.

Authors

Tessa Cressey¹, Kristina Brauburger^{1

2}, Elke Mühlberger³

Affiliations

¹ Department of Microbiology and National Emerging Infectious Diseases Laboratories, Boston University, School of Medicine, Boston, MA, USA.
² Department of Biology, Lund University, Lund, Sweden.
³ Department of Microbiology and National Emerging Infectious Diseases Laboratories, Boston University, School of Medicine, Boston, MA, USA. [email protected].

PMID: 28573612
DOI: 10.1007/978-1-4939-7116-9_6

Abstract

In this chapter, we describe the minigenome system for Ebola virus (EBOV), which reconstitutes EBOV polymerase activity in cells and can be used to model viral genome replication and transcription. This protocol comprises all steps including cell culture, plasmid preparation, transfection, and luciferase reporter assay readout.

Keywords: Ebola virus; Filoviruses; Firefly luciferase assay; Minigenome; Replication; Replication promoter; Reporter gene; Transcription; Zaire ebolavirus; β-galactosidase enzyme assay.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Animals
Ebolavirus / genetics*
Ebolavirus / pathogenicity
Gene Expression Regulation, Viral
Genome, Viral / genetics
Hemorrhagic Fever, Ebola / genetics*
Hemorrhagic Fever, Ebola / virology
Humans
RNA, Viral / genetics
Transcription, Genetic*
Viral Proteins / genetics
Virus Replication / genetics*

Substances

RNA, Viral
Viral Proteins

Abstract

Publication types

MeSH terms

Substances

Grants and funding