Non-replicating vaccinia vector efficiently expresses bacteriophage T7 RNA polymerase

Modified vaccinia virus Ankara (MVA), a host range restricted and highly attenuated vaccinia virus strain, is unable to multiply in human and most other mammalian cell lines. Since viral gene expression is unimpaired in non-permissive cells recombinant MVA viruses are efficient as well as exceptiona...

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Bibliographic Details
Published in:FEBS letters 1995-08, Vol.371 (1), p.9-12
Main Authors: Sutter, Gerd, Ohlmann, Marion, Erfle, Volker
Format: Article
Language:English
Subjects:
Animals
Attenuation
Base Sequence
beta-Galactosidase - biosynthesis
Cells, Cultured
Chick Embryo
Chloramphenicol O-Acetyltransferase - biosynthesis
Chloramphenicol O-Acetyltransferase - genetics
DNA-Directed RNA Polymerases - biosynthesis
DNA-Directed RNA Polymerases - genetics
Expression vector
Fibroblasts
Gene Expression Regulation, Viral - genetics
Genes, Reporter - genetics
Genetic Vectors - genetics
Haplorhini
HeLa Cells
Host restriction
Humans
Molecular Sequence Data
phage T7
Poxvirus
Promoter Regions, Genetic - genetics
Recombinant Fusion Proteins - genetics
T7 RNA polymerase
vaccinia virus
Vaccinia virus - genetics
Vaccinia virus - physiology
Viral Proteins
Virus Replication
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Summary:Modified vaccinia virus Ankara (MVA), a host range restricted and highly attenuated vaccinia virus strain, is unable to multiply in human and most other mammalian cell lines. Since viral gene expression is unimpaired in non-permissive cells recombinant MVA viruses are efficient as well as exceptionally safe expression vectors. We constructed a recombinant MVA that expresses the bacteriophage T7 RNA polymerase and tested its usefulness for transient expression of recombinant genes under the control of a T7 promoter. Using the chloramphenicol acetyltransferase (CAT) gene as a reporter gene, infection with MVA-T7pol allowed efficient synthesis of recombinant enzyme in mammalian cells. Despite the severe host restriction of MVA, enzyme activities induced by infection with MVA-T7pol were similar to those determined after infection with a replication-competent vaccinia-T7pol recombinant virus. Thus, MVA-T7pol may be used as a novel vaccinia vector to achieve T7 RNA polymerase-specific recombinant gene expression in the absence of productive vaccinia virus replication.
ISSN:0014-5793
1873-3468
DOI:10.1016/0014-5793(95)00843-X