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KMID : 1041219960380010036
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Journal of Sericultural and Entomological Science
1996 Volume.38 No. 1 p.36 ~ p.41
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Characterization of Recombinant Baculovirus Ewpressing Polyhedrin Gene of Bombyx mori and Autographa californica Nuclear Polyhedrois Virus
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Abstract
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Recombinant baculoviruses haying expanded host range were selected by coinfection of
Autographa californica NPV and Bombyx mori NPV into Sf-9 and BmN-4 insect cell lines. In order to determine the polyhedra morphology of RecS-A6, one of a recombinant baculovirus, polyhedra of RecS-A6 produced in insect cells were observed by phase contrast microscope and scanning electron microscope. The results revealed that the recombinant baculovirus had a various polyhedra morphology which was different from its parental viruses, suggesting that the various morphology of recombinant baculovirus with an expanded host range was due to the genetic recombinatioH of viral genome. To analyze the genomic recombinantion of the recombinant baculoviruses, genomic DNAs of two parent viruses and RecS-A6 were digested with restriction endonuclease and subjected to agarose gel electrophoresis. Southern blot analysis revealed that RecS-A6 has two polyhedrin gene of AcNPV and BmNPV in a viral genome. Polyhedral protein of recombinant baculovirus was analysed by SDS-PAGE. The result showed that molecular weight of polyhedral protein of RecS-A6 containing two polyhedrin gene of AcNPV and BmNPV was as the 31 kDa band of AcNPV and 30 kDa band of BmNPV.
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