• Korean journal of applied entomology
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• v.28 no.3
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• pp.105-112
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• 1989

The nuclear polyhedrosis viruses of Bombyx mori (BmNPV) and Hyphantria cunea (HcNPV) were characterized by electron microscopic observation, SDS-PAGE of polyhedral and virion proteins, and restriction endonuclease analysis of viral DNAs. Polyhedra of BmNPV were octadecahedral in shape with the diameter of $3 \mu\textrm{m}$, whereas those of HcNPV showed irregular appearances having the diameter of $1.5~2\mu\textrm{m}$. Under alkaline protease inactivated condition, polyhedral proteins of two NPVs were resolved into a major polypeptide, 30~31 KD, and several minor polypeptides by SDS-PAGE. Examination of virion proteins by silver staining after SDS-PAGE showed that BmNPV was composed of 47 polypeptides with M.W. range of 9.6~112 KD and HcNPV was composed of 48 polypeptides with M.W. range of 9.4~111 KD. The approximate genome size of two NPVs were determined by restriction endonuclease analysis: BmNPV and HcNPV were 114.6 Kb, respectively.

• Proceedings of the Korean Society of Sericultural Science Conference
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• 1997.06a
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• pp.73-101
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• 1997

Baculoviruses are characterized by large double-stranded circular DNA genomes and rod-shaped enveloped virions. Bombyx mori nucleopolyhedrovirus(BmNPV) is a major pathogen, which causes severe damage in sericulture. Currently, BmNPV is recogtnized as an improtant tool in molecular biology, especially for expression of useful genes in B.mori cells and silkworm larvae. Our laboratories have focused on the studies of the molecular mechanisms of BmNPV replication and the application of BmNPV to agriculture and medicine. The entire nucleotide sequence of the BmNPV genome has recently determined. The BmNPV genome possessed 135 putative genes and 7 homologous repeated sequence (hrs) regions. Relatively little space, a few to a few hundred base-pairs, was observed between the open reading frames and hrs. Termination codons often overlapped. These results showed a compactly packde BmNPV genome. Based on comparative sequence analyses, we speculated that the ancestor of BmNPV was a baculovirus similar to Autographa californica NPV(AcNPV). The function of the BmNPV genes were characterized by gene deletion analysis; p35 was found to be involved in blocking apoptosis and cysteine proteinase was found to be involved in horizontal virus transmission by degrading viral-infected larval host. By AcNPV and BmNPV coinfection experiments, we identified a BmNPV gene involved in expanding host specificity of AcNPV. The identified gene was likely encoded a DNA helicase based on the amino acid sequence analysis; a few amino acid substitutions in the putative DNA helicase gene resulted in the expansion of host range of AcNPV. These findings indicate that BmNPV evolved within a short period from an AcNPV-like ancestral virus due to rapid evolution including specific amino acid substitutions and gene deletions/insertions.

• Korean Journal of Organic Agriculture
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• v.31 no.4
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• pp.395-412
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• 2023

The morphology and whole genome sequence of Hyphantria cunea nucleopolyhedrovirus W1 (HycuNPV-W1) isolated in Korea were analyzed for the use as an eco-friendly control agent against H. cunea. The HycuNPV-W1 had irregular tetrahedral polyhedra with a size of 1.5-2.2 ㎛ which is similar to that of previously reported HycuNPV isolated in Korea. As a result of whole viral genome analysis, HycuNPV-W1 was composed of 131,353 bp, which is 1,606 bp shorter than that of the previously reported HycuNPV. The G+C content was 45% and six of the homologous repeated regions were found, so there was no significant difference from the previous report. As a result of ORF analysis, HycuNPV-W1 contains total of 145 ORFs which is three ORFs less than the previous report, while two ORFs were exclusively found in HycuNPV-W1. The functions of these ORFs remains unclear and are not considered to have a significant influence on the characteristics of the HycuNPV. The genome vista analysis showed that the overall sequence identity between HycuNPV-W1 and the previously reported HycuNPV was very high. The whole genome of HycuNPV-W1 analyzed was found to be similar to those of the previously reported HycuNPV, however, it is supposed to be a novel resource in Korea with different isolate.

• Korean journal of applied entomology
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• v.61 no.3
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• pp.449-460
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• 2022

The morphology and whole genome sequence of Spodoptera exigua nucleopolyhedrovirus K1 (SeNPV-K1) isolated in Korea were analyzed for the use as an eco-friendly control source against S. exigua. The polyhedra of SeNPV-K1 was amorphous with a size of 0.6-1.8 ㎛, and there was no external difference with the previously reported SeNPV. As a result of analyzing the nucleotide sequence of the whole genome, it was composed of 135,756 bp, which is 145 bp more than that of the previously reported SeNPV. The G+CG+C content was 44% and there were 6 homologous repeated sequences, so there was no significant difference from the previous report. As a result of ORF analysis, SeNPV-K1 had 137, two fewer than those previously reported, and 4 ORFs present only in SeNPV-K1 were confirmed. These 4 ORFs are non-essential genes and were not considered to have a significant influence on the characteristics of the SeNPV. The genome vista analysis showed that the overall sequence similarity between SeNPV-K1 and the previously reported SeNPV was very high. The whole genome of SeNPV-K1 analyzed for the first time in Korea was found to be similar to the previously reported SeNPV, but it was confirmed that it was a novel resource in Korea with different isolate.

• Journal of Sericultural and Entomological Science
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• v.38 no.1
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• pp.36-41
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• 1996

Recombinant baculoviruses having expanded host range were selected by coinfection of Autographa california NPV and Bombyx mori NPV into Sf-9 and BmN-4 insect cell lines. In order to determine the polyhedra morhplogy 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 morhpology of recombinant baculovirus with an expanded host range was due to the genetic recombination 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.

• 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.

• International Journal of Industrial Entomology and Biomaterials
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• v.3 no.1
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• pp.13-18
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• 2001

The role of polyhedrin in the polyhedra production in baculovirus Autograha californica Nucelopolyhedro-sisvirus (AcNPV) was studied by over-expression of AcNPV polyhedrin or heterologous polyhedrin from Bombyx mori (Bm) NPV or Spodoptera exigua (Se) NPV. The transfer vectors containing additional polyhedrin from AcNPV, BmNPV, or SeNPV were constructed and cotransfected with bacmid bApGOZA into Sf9 cells. The resulting recombinants, designated as vApAcPol, vApBmPol, and vApSePol were tonstructed, and the polyhedra production of the recombinant was characterized. All of the recombinants produced polyhedra in the nucleus, and the polyhedrin was over-expressed. Among three recombinants, vApAcPol and vApBmPol were discriminated by their larger polyhedra size than that of wild type AcNPV, and vApSePol also produced larger polyhedra than wild type SeNPV polyhedra.

• Journal of Sericultural and Entomological Science
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• v.38 no.1
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• pp.42-47
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• 1996

To investigate the host range determining factors of nuclear polyhedrois virus (NPV), Autographa california 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 expanded host 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-SacI 0.6 kb DNA fragments of RecS-A6 and RecB-727 were cloned and sequenced. The results showed that of wild type BmNPV helicase gene, suggesting that the expanded host range of recombinant baculoviruses was due to the insertion of BmNPV helicase gene into AcNPV viral genome.

• International Journal of Industrial Entomology and Biomaterials
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• v.3 no.1
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• pp.37-41
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• 2001

The ecdysteroid UDP-glucosyltransferase (egt) gene isolated from Bombyx mori nuclear polyhedrosis virus (BmNPV) K1 strain was compared to its homologue from Autographa californica NPV (AcNPV) and Bm NPV T3. The egt gene of BmNPV-K1 encoded 506 amino acid open reading frame, and was 99.6% identical at the amino acid level and 99.2% identical at the nucleotide level to BmNPV T3. The BmNPV-K1 egt gene showed highly identity to AcNPV and BmNPV T3 strain. The BmNPV-K1 egt gene was different from amino acid sequence at 2 positions, 19 and 72, in BmNPV T3. The genomic location of egt gene in the BmNPV-K1 was confirmed by Southern blot analysis and its expression patterns at the transcriptional level in the infected cells were confirmed by Northern hybridization analysis. Transcripts of the egt of Bm NPV-K1 peaked around 12 hrs postinfection (p.i.) and reduced at 24 hrs p.i.

• Korean journal of applied entomology
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• v.28 no.1
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• pp.10-15
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• 1989

This studies were carried out to selected high pathogenic nuclear polyhedrosis viruses(NPVs) against Pseudaletia(=Leucania) separata for the introduction of microbiol control of the insect NPV in Korea. Among 21 NPVs, Sesamia inferens and 4 P. separata NPV strains were highly pathogenic against P. separata when fed orchard grass leaves smeared virus suspension on the 2nd instar larvae. Three NPV strains (LsNPV-F, LsNPV-G, LsNPV-Y) were more susceptible to the younger instar than the older instar P. separata larvae when fed artificial diet mixed with the virus to the insect.