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PubMed · 1629707

Foreign gene expression by a baculovirus vector with an expanded host range.

H Mori·H Nakazawa·N Shirai·N Shibata·M Sumida·F Matsubara
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Abstract

A nuclear polyhedrosis virus (NPV) (Baculoviridae)-based gene expression system was improved by DNA recombination. The BmN cell line established from Bombyx mori and the Sf21 cell line established from Spodoptera frugiperda are non-permissive for Autographa californica multicapsid NPV (AcMNPV) and B. mori NPV (BmNPV) replication, respectively. After cotransfection of AcMNPV DNA and BamHI-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, B. mori. 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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BaculoviridaeCell LineGene ExpressionGenetic VectorsRecombination, GeneticJournal Article

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BibTeXRIS

H Mori, H Nakazawa, N Shirai, N Shibata, M Sumida, F Matsubara. 1992. Foreign gene expression by a baculovirus vector with an expanded host range.. https://doi.org/10.1099/0022-1317-73-7-1877

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