A nuclear polyhedrosis virus (NPV) ()-based gene expression system was improved by DNA recombination. The BmN cell line established from and the Sf21 cell line established from are non-permissive for multicapsid NPV (AcMNPV) and NPV (BmNPV) replication, respectively. After cotransfection of AcMNPV DNA and HI-digested BmNPV DNA into Sf21 cells, progeny viruses were isolated by plaque purification on BmN cell monolayers and the host specificity of one viral isolate was analysed. The virus had a wider host range, and replicated and produced polyhedra in Sf21 cells, BmN cells and larvae of the silkworm, DNA restriction endonuclease analysis showed that the isolate was a hybrid of AcMNPV and BmNPV. Using the AcMNPV transfer vector pAcYM1 a portion of the polyhedrin gene of the hybrid virus was replaced with the coding region of the firefly luciferase gene, producing a recombinant virus. The latter expressed firefly luciferase in both cell lines and in silkworm larvae under the control of the polyhedrin promoter.


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