• Korean journal of applied entomology
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• v.27 no.4
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• pp.211-218
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• 1988

Polyhedral proteins and the endogenous alkaline protease associated with larval-derived polyhedra of nuclear viruses isolated from Spodoptera litura, Bombyx mori, and Hyphantria cunea were investigated. Polyhedral proteins prepared under alkaline protease heat-inactivated condition were separated as one band with 31Kd in S. litura a H. cunea NpV and 30Kd in B. mori NPV by the SDS-polyacrylamide gel electroptoresis. Whereas polyhedral proteins without heat-inactivation were degraded into smaller polypeptides with a certain pattern in alkaline solution. The results of double-immunodiffusion and western blot analysis with antisera against polyhedral proteins indicated that those three polyhedral proteins had common antigenic determinants and the degradation of polyhedral proteins by alkaline protease could be confirmed.

• Proceedings of the Korean Society of Applied Entomolohy Conference
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• 2001.05a
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• pp.139-139
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• 2001

• Proceedings of the Korean Society of Applied Entomolohy Conference
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• 2001.11a
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• pp.157-157
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• 2001

• Proceedings of the Korean Society of Applied Entomolohy Conference
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• 2001.11a
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• pp.156-156
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• 2001

• Korean journal of applied entomology
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• v.34 no.3
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• pp.218-223
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• 1995

A nuclear polyhedrosis virus isolated from the oriental tobacco budworm larvae, Helicoverpa assulta (Guenee) was characterized by electron microscopy, SDS-PAGE, restriction endonuclease analysis and cross infectivity. The shape of a polyhedron was $1.0\mu\textrm{m}$ in average with icosahdral outline, and the virus particle was $65nm\times300nm$ in average with rod-shape. The nuclear polyhedrosis virus was contained a single nucleocapsid within a viral envelope embedded in a polyhedron. The polyhedral protein was composed of a single polypeptide with a M.W. of 31 Kd. The genome size of the virus by restriction endonuclease analysis was about 120 Kb. Among several nuclear polyhedrosis viruses, the nuclear polyhedrosis virus from Helicoverpa assulta (HaNPV) and Autographa california nuclear polyhedrosis virus (AcNPV) were infected the oriental tobacco budworm larvae.

• Korean journal of applied entomology
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• v.38 no.2
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• pp.139-144
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• 1999

We have now constructed a novel recombinant baculovirus producing fusion protein with Autogrqha c.uliforrzica nuclear polyhedrosis virus (AcNPV) polyhedrin and green fluorescent protein (GFP). The fusion protein expressed by the recombinant baculovirus in insect cells was characterized. The GFP gene was introduced under the control of polyhedrin gene promoter of AcNPV, by fusion in the front or back of intact polyhedrin gene. The recombinant baculoviruses were named as Ac-GFPPOL or Ac-POLGFP. respectively. As expected, the 56 kDa fusion protein was expressed in the recombinant virus-infected cells. Interestingly. however, the fluorescence of GFP in the cells infected with Ac- POLGFP was only detected within the nuclei. and that was observed as polyhedra-like granular particles. In the microscopy of cells infected with Ac-GFPPOL, furthermore, GFP was detected in both cytoplasm and nuclei although most of GFP were present within the nuclei. However, fusion protein produced by recombinant virus did not form polyhedra although the fusion protein was fused with polyhedrin and GFP. It is suggested that difference of GFP location in the infected cells appear to be involved in the region of polyhedrin in the fusion protein, and the polyhedrin in the fusion protein might be responsible for the polyhedra-like granular particles present within nuclei.

• Korean journal of applied entomology
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• v.36 no.4
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• pp.341-350
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• 1997

We have now constructed a novel recombinant baculovirus producing fusion protein with Autographa californica nuclear polyhedrosis virus (AcNPV) polyhedrin and Bacillus thuringiensis(Bt) cryIA(c) crystal protein. The fusion protein expressed by the recombinant baculovirus in insect cells was characterized. The N-terminal of cryIA(c) gene of Bt subsp. kurstaki HD-73 was introduced under the control of polyhedrin gene promoter of AcNPV, by fusion in the front of intact polyhedrin gene or by insertion into the HindIII site in polyhedrin gene. The recombinant baculoviruses were named as BtrusI or BtrusII, respectively. Although single transcript from the fusion protein gene was apparently observed. BtrusI was produced the two proteins, 92 kDa fusion protein and only polyhedrin. In addition, fusion protein produced by BtrusI did not form polyhedra. Interestingly, however, the cells infected with BtrusII did not show a 33 kDa polyhedrin band as a cells infected with BtrusI. Cells infected with BtrusII were only produced fusion protein, but the polyhedra formed by fusion protein was not observed. To determine the insecticidal toxicity of fusion protein, therefore, Sf9 cells infected with BtrusI were inoculated to Bombyx mori larvae. Sf9 cells infected with BtrusI that expressed the fusion protein caused larval mortality although the insecticidal toxicity was low. In conclusion, our results clearly demonstrated that the fusion protein with polyhedrin and Bt cryIA(c) crystal protein have a insecticida toxicity.

• Korean journal of applied entomology
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• v.29 no.3
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• pp.184-189
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• 1990

A nuclear polyhedrosis virus (NPV) of the tobacco cutworm, Spodoptera litura would be a promisible agent for the control of the insect. To develop a viral insecticide using S. litura NPV, effect of spray on soybean leaves, temperature, storage, an sunlight on the pathogenicity of the virus were studies as follows: Median lethal concentration ($LC_{50}$) of the virus sprayed on the leaves against the third and the fifth instar larvae were $1.301\times10^{4 PIBS}/ml$ and $1.087\times10^{5 PIBS}/ml$, respectively. On the concentration of $1.0\times10^{5 PIBS}/ml$, median lethal times ($LT_{50}$) were 7.3 days for the 3rd and 8.9 days for the 5th instar larvae. Stability of S. litura NPV was quickly decreased at the higher temperate than $60^{\circ}C$ and at the longer exposure to the higher temperature. Storage of the virus at $-20^{\circ}C$ was kept higher pathogenicity than $4^{\circ}C$ and $25^{\circ}C$. Viral activity was maintained more than 10 days in the sprayed-under leaves, but decreased at 3 day after spray in th sprayed-on the leaf surface when exposed the virus to sunlight.

• Microbiology and Biotechnology Letters
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• v.21 no.6
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• pp.513-519
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• 1993

The alkaline protease in the polyhedra preparation of Spodoptera litura nuclear polyhedrosis virus was successfully inactivated by heating at 100C for 20 minutes. SDS-PAGE analysis indicated that heat inactivated polyhedra is composed of major proteins of 31kDa and presumptive its polymer protein of 62kDa. However, this polyhedra was converted into several smaller molecular weight proteins when treated with midgut juice, but not by treatment with heat-inactivated midgut juice.

• Korean journal of applied entomology
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• v.30 no.2
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• pp.144-149
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• 1991

Biochemical characteristics of Spodoptera uigua nuclear polyhedrosis virus (SeNPV) isolated in Jinju were studied. SeNPV contained a number of nucleocapsids within a viral envelope embeded in polyhedra. The polyhedral protein of SeNPV was composed of a single polypeptide with a M. W. of 30kd. Double-immunodiffusion test showed that the polyhedral protein of SeNPV had common antigenic determinants with SINPV and BmNPV. Virion proteins of SeNPV were resolved into 49 polypeptides by silver staining after SDS-PAGE. The approximate genome size of SeNPV by restriction endonuclease analysis was 1l0kb.