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

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...

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Bibliographic Details
Published in:Journal of general virology 1992-07, Vol.73 (7), p.1877-1880
Main Authors: Mori, H, Shirai, N, Shibata, N, Sumida, M, Matsubara, F
Format: Article
Language:English
Subjects:
Autographa californica
Baculoviridae
Baculoviridae - genetics
baculovirus
Biological and medical sciences
Biotechnology
Bombyx mori
cell culture
Cell Line
Fundamental and applied biological sciences. Psychology
Gene Expression
Genetic engineering
Genetic technics
Genetic Vectors
Methods. Procedures. Technologies
nuclear polyhedrosis virus
Recombination, Genetic
Spodoptera frugiperda
Vectors (cloning, transfer, expression). Insertion sequences and transposons
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Summary: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.
ISSN:0022-1317
1465-2099
DOI:10.1099/0022-1317-73-7-1877