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PMID: 8986823 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Rescue of a segmented negative-strand RNA virus entirely from cloned complementary DNAs.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 93 ·No. 26 ·1996-12-24 ·Pages 15400-4

Bridgen A, Elliott RM

Abstract

We provide the first report, to our knowledge, of a helper-independent system for rescuing a segmented, negative-strand RNA genome virus entirely from cloned cDNAs. Plasmids were constructed containing full-length cDNA copies of the three Bunyamwera bunyavirus RNA genome segments flanked by bacteriophage T7 promoter and hepatitis delta virus ribozyme sequences. When cells expressing both bacteriophage T7 RNA polymerase and recombinant Bunyamwera bunyavirus proteins were transfected with these plasmids, full-length antigenome RNAs were transcribed intracellularly, and these in turn were replicated and packaged into infectious bunyavirus particles. The resulting progeny virus contained specific genetic tags characteristic of the parental cDNA clones. Reassortant viruses containing two genome segments of Bunyamwera bunyavirus and one segment of Maguari bunyavirus were also produced following transfection of appropriate plasmids. This accomplishment will allow the full application of recombinant DNA technology to manipulate the bunyavirus genome.

MeSH Terms
Base Sequence Bunyamwera virus/genetics,metabolism Cloning, Molecular DNA Primers DNA, Complementary HeLa Cells Humans Molecular Sequence Data Plasmids Polymerase Chain Reaction RNA Viruses/genetics,metabolism RNA, Viral/metabolism Transcription, Genetic Transfection Viral Proteins/biosynthesis
Chemicals
DNA Primers DNA, Complementary RNA, Viral Viral Proteins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Bridgen A
Institute of Virology, University of Glasgow, Scotland.
Elliott R M
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.93.26.15400
Published
1996-12-24
Pages
15400-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC26416
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.93.26.15400 PMC Full Text View journal details
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