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 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.
Sutter
G
G
Institut für Molekulare Virologie, GSF-Forschungszentrum für Unwelt und Gesundheit GmbH, Oberschleissheim, FRG.
Ohlmann
M
M
Erfle
V
V
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D002642:Chick Embryo; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000096:biosynthesis* / Q000235:genetics; D005347:Fibroblasts; D015967:Gene Expression Regulation, Viral / Q000235:genetics*; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D000882:Haplorhini; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014616:Vaccinia virus / Q000235:genetics* / Q000502:physiology; D014764:Viral Proteins; D014779:Virus Replication; D001616:beta-Galactosidase / Q000096:biosynthesis
D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D015500:Chloramphenicol O-Acetyltransferase; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; D001616:beta-Galactosidase