Sutter G, Ohlmann M, Erfle V
Institut für Molekulare Virologie, GSF-Forschungszentrum für Unwelt und Gesundheit GmbH, Oberschleissheim, FRG.
FEBS Lett. 1995 Aug 28;371(1):9-12. doi: 10.1016/0014-5793(95)00843-x.
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.
安卡拉痘苗病毒（MVA）是一种宿主范围受限且高度减毒的痘苗病毒株，无法在人类及大多数其他哺乳动物细胞系中增殖。由于病毒基因表达在非允许细胞中不受影响，重组MVA病毒是高效且极其安全的表达载体。我们构建了一种表达噬菌体T7 RNA聚合酶的重组MVA，并测试了其在T7启动子控制下用于重组基因瞬时表达的效用。以氯霉素乙酰转移酶（CAT）基因作为报告基因，用MVA-T7pol感染可使哺乳动物细胞高效合成重组酶。尽管MVA对宿主有严格限制，但用MVA-T7pol感染诱导的酶活性与用具有复制能力的痘苗-T7pol重组病毒感染后测定的酶活性相似。因此，MVA-T7pol可作为一种新型痘苗载体，在无痘苗病毒有效复制的情况下实现T7 RNA聚合酶特异性重组基因表达。
