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KMID : 1041219960380010042
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Journal of Sericultural and Entomological Science
1996 Volume.38 No. 1 p.42 ~ p.47
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Molecular Biological Characterization of Recombinant Baculovirus with an Expanded Host Range
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Abstract
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To investigate the host range determining factors of nuclear polyhedrosis virus (NPV),Autographa californica NPV and Bombyx mori NPV were coinfected into the two different cell lines, BmN-4 and sf-9. The recombinant baculoviruses, RecS-A6 and RecB-727 which have an expanded host range, were isolated from Sf-9 and BmN-4 cell lines, respectively. The molecular biological characteristics of the recombinant baculoviruses were investigated. The pathogenicity of RecB-727 was similar to that of wild type BmNPV, while the pathogenicity of RecS-A6 was relatively lower than that of wild type BmNPV. The restriction enzyme digestion patterns of parental viruses and recombinant viruses showed that the recombinant virus has an expandedhost range by genetic recombination. Southern blot analysis revealed that the p10 gene of RecB-727 was derived from AcNPV genomic DNA, while RecS-A6 has p10 gene of BmNPV in a viral genome. To investigate the host range expansion mechanism of recombinant baculovirus, HindIII-Sacl 0.6 kb DNA fragments of RecS-A6 and RecB-727 were cloned and sequenced. The results showed that nucleotide sequence in 0.6 kb fragment of recombinant baculoviruses were identical to that of wild type BmNPV helicase gene, suggesting that the expanded host range of recombinant baculoviruses was due to the insertion of BmNPV helicasegene into AcNPV viral genome.
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