Rescue of measles virus using a replication-deficient vaccinia-T7 vector

A system which allows the reconstitution of measles virus (MV) from cloned cDNA is described. The severely host cell restricted vaccinia vector MVA-T7 expressing bacteriophage T7 RNA polymerase was used to generate full-length antigenomic MV RNA and simultaneously the mRNAs encoding the viral N, P a...

Full description

Saved in:
Bibliographic Details
Published in:Journal of virological methods 1997-02, Vol.64 (1), p.57-64
Main Authors: Schneider, Henriette, Spielhofer, Pius, Kaelin, Karin, Dötsch, Christina, Radecke, Frank, Sutter, Gerd, Billeter, Martin A.
Format: Article
Language:English
Subjects:
Animals
Bacteriophage T7 - enzymology
Base Sequence
Biological and medical sciences
Cell Line
Cercopithecus aethiops
Defective interfering RNAs
Defective Viruses - genetics
DNA, Viral - analysis
DNA-Directed RNA Polymerases - genetics
Fundamental and applied biological sciences. Psychology
Genetic Vectors
Genome, Viral
HeLa Cells
Humans
Measles
Measles virus - genetics
Measles virus - isolation & purification
Measles virus - physiology
Microbiology
Molecular Sequence Data
MVA-T7
Techniques used in virology
Vaccinia virus - genetics
Vero Cells
Viral Proteins
Virology
Virus Replication
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:A system which allows the reconstitution of measles virus (MV) from cloned cDNA is described. The severely host cell restricted vaccinia vector MVA-T7 expressing bacteriophage T7 RNA polymerase was used to generate full-length antigenomic MV RNA and simultaneously the mRNAs encoding the viral N, P and L proteins in order to produce replicationally and transcriptionally active nucleocapsids. The functionality of the N, P and L proteins was demonstrated first by their ability to rescue MV specific subgenomic RNAs. Assembly and budding of reconstituted MV was shown by syncytia formation and subsequently by virus isolation. The inability of MVA-T7 to produce progeny virus in most mammalian cells circumvents the necessity to separate the reconstituted MV from the MVA-T7 helper virus. Since all components are expressed transiently, this system is especially suitable for studying the functions of N, P and L. Furthermore, it is useful for investigating later steps in the MV life cycle.
ISSN:0166-0934
1879-0984
DOI:10.1016/S0166-0934(96)02137-4