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Non‐replicating vaccinia vector efficiently expresses bacteriophage T7 RNA polymerase
Author(s) -
Sutter Gerd,
Ohlmann Marion,
Erfle Volker
Publication year - 1995
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)00843-x
Subject(s) - vaccinia , bacteriophage , virology , polymerase , t7 rna polymerase , vector (molecular biology) , rna polymerase , biology , dna , rna , genetics , gene , recombinant dna , escherichia coli
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.