• Microbiology and Biotechnology Letters
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• v.26 no.2
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• pp.161-166
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• 1998

To study the usefulness of Se301 cells, which is originated from Spodoptera exigua and has susceptibility to the Autographa californica NPV (AcNPV), as a host for the AcNPV-based expression vector system, we compared the characteristics of AcNPV in Se301 and Sf-21 cells. The symptom by viral infection was similar in both of cells, but the ratio of polyhedra released from the cell was higher in Se301 cells than in Sf-21 cells. The overall PIB productivity of AcNPV was similar in both cells but the size of polyhedra was larger in Se301 cells. While the polyhedrin expression efficiency was about 2.4 times higher in Se301 cells than in Sf-21 cells, the viral growth was higher in Sf-21 cells. These results suggested that Se301 cell is very useful in the AcNPV-based expression system as a host.

• International Journal of Industrial Entomology and Biomaterials
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• v.22 no.2
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• pp.75-82
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• 2011

To elucidate the novelty of Hyphantria cunea nucleopolyhedrovirus (HcNPV) isolated in Korea, polyhedrin and inhibitor of apoptosis (iap) gene structures were determined and analyzed. The analysis of HcNPV polyhedrin showed a little difference with 97.6% at the nucleotide level but no difference at the amino acid level when compared with that of previously reported H. cunea NPV (HycuNPV). On the other hand, iap genes showed variable differences with 89.0-99.6% nucleotide and 90.0-99.6% amino acid sequence identities. Especially, the 5' and 3' non-coding flanking sequences of iap1 gene had lower degree of identity with those of HycuNPV. Although the phylogenetic analyses using polyhedrin and iap genes showed that HcNPV is closely related with HycuNPV, we could provide that HcNPV is a novel isolate having novel gene structures.

• Journal of Microbiology and Biotechnology
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• v.8 no.6
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• pp.685-691
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• 1998

Baculovirus Hyphantrin. cunea nuclear polyhedrosis virus (HcNPV) insecticide containing the insecticidal protein (ICP) gene from Bacillus thuringiensis subsp. kurstaki HD1 was constructed using a lacZ-HcNPV system. The ICP ($\delta$-endotoxin) gene was placed under the control of the polyhedrin gene promoter of the HcNPV. A polyhedrin-negative virus was derived and named ICP-HcNPV insecticide. Then, the insertion of the ICP gene in the ICP-HcNPV genome was confirmed by Southern hybridization analysis. Polyacrylamide gel electrophoresis (PAGE) analysis of the Spodoptera frugiperda cell extracts infected with the ICP-HcNPV showed that the ICP was expressed in the insect cells as 130 kDa at 5 days post-infection. The ICP produced in the cells was present in aggregates. When extracts from the cells infected with the ICP-HcNPV were fed to 20 Bombyx mori larvae, the following mortality rate was seen; 8 larvae at 1 h, 10 larvae at 3 h, and 20 larvae at 12 h. These data indicate that the B. thuringiensis ICP gene was expressed by the baculovirus insecticide in insect cells and there was a high insecticidal activity. The biological activities of the recombinant virus ICP-HcNPV were assessed in conventional bioassay tests by feeding virus particles and ICP to the insect larvae. The initial baculovirus insecticide ICP-HcNPV was developed in our laboratory and the significance of the genetically engineered virus insecticides is discussed.

• Microbiology and Biotechnology Letters
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• v.18 no.6
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• pp.660-661
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• 1990

A stable preservation method of extracellular non-occluded virion of Autographa californica nuclear polyhedrosis virus (AcNPV) was studied. AcNPVL-1 strain infected to Spodoptera frugiperda cell line and then the culture media were centrifuged. After centrifugation the supernatant containing extracellular nonoccluded virions of the AcNPV was harvested and incubated at $4^{\circ}C$ . Even after the extracellular nonoccluded virions were incubated at $4^{\circ}C$ for about 11 years, the infectivity and multiplication property of the nonoccluded virions in the S. frugiperda cell line were normal. However the titers of the nonoccluded virions in TC-100 medium measured about 11 years ago decreased from $8.9 \times 10^7\; to \;3.8 \times 10^5$ pfu per ml. The AcNPV genome DNA fragment patterns from digestion with Hind11 and EcoRI restriction endonucleases did not change. The AcNPV nonoccluded virions were stable at $4^{\circ}C$ in the cultured medium of more than 10 years and the preservation of AcNPV nonoccluded virions at $4^{\circ}C$ is easy and useful for handling.

• Journal of Sericultural and Entomological Science
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• v.41 no.2
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• pp.102-107
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• 1999

To analysis a promoter strength of Atographa californica nucler polyhedrosis virus (AcNPV) IE1 gene, an immediate viral gene, ${\beta}$-glactosidase gene as a reporter gene was introduced under the control of the IE1 promoter. The restriction fragment containing IE1 promoter and ${\beta}$-galctosidase gene from pAcIE1-gal were inserter into pBacPAK9 to yield transfer vector pAcNPV-IE1-gal. The pAcNPV-IE1-gal was cotransfected with AcNPV genomic DNA BacPAK6 into Sf9 cells to produce recombinant baculovirus AcNPV-IE1-gal. In addition, recombinant bacvulovirus AcNPV-gal, which express ${\beta}$-galac-tosidase under the control of the polyhedrin promoter, was constrer, was constructed to compared with AcNPV-IE1-gal. The recombinant viruses were respectively infected into Sf9 cells and characterized by the virus titer and expression of ${\beta}$-galactoxidase in Sf9 cells. The promoter strength of IE1 and polyhedrin promoters was determined by the amount of ${\beta}$-galactosidase secreted into medium by viral infection. The titer of AcNPV-IE1-Gal determined by plaque assays in Sf9 cells was similar to that of AcNPV-gal. However, expression level of ${\beta}$-galactosidase by AcNPV-IE1-gal was significantly lower than that by AcNPV-gal. In conclusion, promoter strength of IE1 was approximately 25-fold lower than that of polyhedrin.

• Applied Microscopy
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• v.21 no.1
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• pp.21-26
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• 1991

Midgut cells of the dead Dendrolimus spectabilis larvae by infection of D. spectabilis nuclear polyhedrosis virus (DsNPV) were observed by transmission electron microscopy. DsNPV replicated in the nucleus of the infected midgut cells and the virogenic stroma of DsNPV appeared in the nucleus, from which nucleocapsids were formed. Also the formation of polyhedral inclusion bodies were observed in the nucleus.

• International Journal of Industrial Entomology and Biomaterials
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• v.3 no.1
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• pp.25-29
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• 2001

We have cloned and characterized an antiapoptotic gene, p35, which blocks apoptosis, from Bombyx mori nuclear polyhedrosis virus (BmNPV) K1 strain. The 897 bp p35 has an open reading frame of 299 amino acids. The BmNPV-K1 p35 showed a high identity to Autographa californica nuclear polyhedrosis virus and BmNPV T3 strain. The BmNPV-K1 p35 was different from the amino acid sequences of BmNPV T3 at 6 positions. The p35 gene of BmNPV-K1 was 99.2% identical at the nucleotide level and 98% identical at the amino acid level to BmNPV T3. The location of p35 gene in the BmNPV-K1 genome was confirmed by Southern blot analysis and its expression patterns at the transcriptional level in the infected cells were con- firmed by Northern hybridization analysis.

• Journal of Sericultural and Entomological Science
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• v.38 no.2
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• pp.144-149
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• 1996

To develope the baculovirus expression vector system (BEVS) using Spodoptera exigua nuclear polyhedrosis virus (SeNPV), we characterized the polyhedrin of SeNPV. The SeNPV polyhedra was irregular and composed of the major protein molecular weight of 30 kDa determined by electronmicroscopy and SDS-AGE analysis, respectively. The nucleotid suquences of 876 bases including the coding region of polyhedrin gene was determined and it was revealed that the polyhedrin gene is located within Xho I 3.0Kb and Nco I 6.0 Kb by Southern blot analysis, respectively. Also, the Xho I 3.0 Kb and the Nco I 6.0 Kb fragments were cloned and restriction enzyme map of these clones were determined.

• Journal of Sericultural and Entomological Science
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• v.40 no.2
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• pp.105-110
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• 1998

The baculovirus egt gene encodes an ecdysteroid UDP-glucosyltransferase(EGT) which catalyzes the transfer of glucose from UDP-glucose to the insect moltion hormone ecdysteroid resulting in a functionally inactive ecdysteroid. In baculovirus-infected insect larvae, EGT has been shown block molting and pupation. In this study, we compared the development of 4th and 5th instar silkworm, Bombyx mori, larvae injected with either wild-type bombyx mori nucleopolyhedrovirus (BmNPV) or a mutant BmNPV(BmEGTZ) in which the egt gene was disrupted by the insertion of a lacZ gene cassette. Larvae injected with BmEGTZ died roughly 12 h more rapidly compared to indentical larvae infected with BmNPV. In addition, BmEGTZ- infected larvae prematurely stopped feeding and gain less weight compared to BmNPV-infected larvae. In order to investigate why BmEGTZ-infected larvae died more rapidly than BmNPV-infected larvae, the array of hemolymph proteins in BmEGTZ-or BmNPV-infected larvae were analyzed by SDS-PAGE. The hemolymph of BmEGTZ-infected larvae showed virus-specific proteins, including polyhedrin, about 12 h earlier than the hemolymph of BmNPV-infected larvae

• Korean journal of applied entomology
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• v.32 no.4
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• pp.407-413
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• 1993

Twelve recombinant viruses with wider host range were plaque purified after coinfectian of Autographa cahjornica and Bombyx mOT! NPVs into Sf9 ar BmN-4 cells. Restriction endonucleases analysis of the recombinant's DNAs showed that the recombinatIOn between AcNPV and BmNPV genomes had occurred more than once. When the recombinam RecB-8, derived from BrnN-4 cells, was observed by electron rntcroscopy, the shape of the polyhedron was a regular tetrahedron, and few virions were occluded into a polyhedron.