• International Journal of Industrial Entomology and Biomaterials
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• 제6권2호
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• pp.179-181
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• 2003

$F(ab`)_2$-ELISA and direct antigen coating-ELISA (DAC-ELISA) were evaluated in the detection of purified Bombyx mori nuclear polyhedrosis virus (BmNPV) and nuclear polyhedrosis virus infection in silkworm larvae inoculated with BmNPV polyhedra. Although nanogram levels of BmNPV was detected in both DAC- and $F(ab`)_2$-ELISA, similar concentrations of antigen was detected in case of F(ab’)$_2$-ELISA even at higher dilution of antibody (up to 1 : 20 K). One hundred percent nuclear polyhedrosis infection was detected 6 hrs after inoculation in BmNPV infected silkworm larvae by $F(ab`)_2$-ELISA. On the other hand, detection of 100% infection was observed only three days after inoculation in DAC-ELISA. In this study, it was observed $F(ab`)_2$-ELISA was more sensitive than DAC-ELISA in the detection of purified BmNPV as well as nuclear polyhedrosis infection in silkworm larvae.

• 한국응용곤충학회지
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• 제34권3호
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• pp.218-223
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• 1995

담배와 고추의 주요해충인 담배나방 [Helicoverpa assulta(Guenee)]의 미생물적 방제법을 개발하기 위하여, 아직 국내에서 보고되지 않은 담배나방 핵다각체병 바이러스 담배나방 사충으로 부터 분리하고, 바이러스의 전자현미경 관찰 및 바이러스 DNA의 제한효소 분석 등을 통한 생화학적 특성을 조사하였다. 담배나방 핵다각체병 바이러스의 다각체는 구형에 가까운 20면체로서 크기는 평균 약 1.0$\mu$m 정도이고, 다수의 바이러스 입자가 다각체 단백질에 포매되어 있었다. 포매된 바이러스 입자는 한개의 봉상 nucleocapsid가 하나의 envelope 내에 매립된 형태의 SNPV(single embedded nuclear polyhedrosis virus)로, 형태는 전형적인 봉상으로서 그 크기는 약 $65nm\times300nm$ 였다. 또한 다각체 단백질의 SDS-PAGE 분석 결고, 분자량은 약 31 Kd이었으며, 담배나방 핵다각체병바이러스 DNA를 분리하여 EcoRI. HindIII 등 수종의 제한효소로 처리분석한 결과, 그 genome의 크기는 약 120Kb 정도였다. 본 핵다각체병바이러스는 담배나방 유충에만 병원성을 나타내 기주특이성을 보였다.

• 한국잠사곤충학회지
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• 11호
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• pp.59-62
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• 1970

1. 한국에서 작잠농병을 발생시키는 주된 Virus는 핵질다색체 Virus 이다. 2. Virus 묶음은 봉입체단백직의 분자구조내에 함입되어있다. 3. 봉입체단백질에 존재하는 Virus묶음은 평균4개의 간상형 Virus입자로 되어있으며 이를 싸고있는 막은 두개로 되어있다. 4. 봉입체단백질내에 존재하는 Virus묶음은 질서와 균형있게 배열된 것이 아니고 되는데로 산재해 있는 것 같다. 5. Virus입자와 봉입예단백질은 전염된 세포의 염색체에서 형성된 소위 "Virogenic stroma"에서 생겨진것이다.

• Applied Microscopy
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• 제21권1호
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• pp.21-26
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• 1991

Dendrolimus spectabilis nuclear polyhedrosis virus(DsNPV)에 감염되어 죽은 솔나방(D. spectabilis)유충의 중장세포를 전자현미경(TEM)으로 관찰하였다. 감염된 중장세포의 핵에서 DsNPV는 복제하였으며, DsNPV의 virogentic stroma가 핵속에 나타났고, nucleocapsid도 형성되었다. 또한 NPV의 polyhedra형성을 핵에서 관찰했다.

• 한국미생물·생명공학회지
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• 제21권6호
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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.

• International Journal of Industrial Entomology and Biomaterials
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• 제1권1호
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• pp.29-33
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• 2000

We cloned and characterized a very late expression factor 1 gene, vlf-1, which regulates the level of very late gene transcripts, from Bombyx mori nuclear polyhedrosis virus (BmNPV) K1 strain. The 1,140 bp vlf-1 has an open reading frame of 379 amino acid and a MW of 44 kDa. The vlf-1 nucleotide sequence of BmNPV-Kl showed high homology with Autographa californica nuclear polyhedrosis virus and BmNPV T3 strain so far known, and its deduced amino acid residues were identical to those of BmNPV T3. The location of vlf-1 in the BmNPV-Kl genome was confirmed by Southern blot analysis and its expression patterns at the transcriptional level were confirmed by Northern hybridization analysis.

• 한국미생물·생명공학회지
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• 제21권5호
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• pp.406-413
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• 1993

This study was performed to produce monoclonal antibodies to the major polyhedral inclusion body (PIB) antigen of the occluded form of Hyphantria cunea nuclear polyhedrosis virus (HcNPV). PIB proteinS were purfied from the Spodoptera frugiperda cell infected with HcNPVs. Using the purified PIB protein, BALB/C mice were immunized 3 times with 2 weeks intervals. The spleen were removed and fused with Sp2/0-Ag14, mouse myeloma cells.

• 한국잠사곤충학회지
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• 제40권2호
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• pp.111-116
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• 1998

To construct transformed Bm5 cells, Autographa californica nuclear polyhedrosis virus (AcNPV)IE1 gene, an immediate early viral gene was firstly used in this study. AcNPV IE1 gene, which shares on 95.3% uncleotide sequence homology with Bombyx mori nuclear polyhedrosis virus (BmNPV) IE1 gene, was isolated and cloned into pBluescript. Neomycin gene from pKO-neo was inserted under the control of the IE1 promoter to yield pAcIE1-neo. The plasmid pAcIE1-neo was transfected into Bm5 or Sf9 cells, and neomycin-resistant cells were selected in TC100 medium containing 10% fetal bovine serum (FBS) and 1 mg/$m\ell$ G418 for two weeks. Individual clones were picked and each was amplified for further characterization. The genomic DNA from neomycin-resistnt cells was isolated and characterized by PCR using AcNPV IE1 gene-specific primers and by Southern blot analysis using neomycin gene probe. We concluded that AcNPV IE1 gene was functional in B. moridrived Bm5 cells as well as Spodaptera frugiperda-derived Sf9 cells to produce stably-transformed insect cells.

• 한국잠사곤충학회지
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• 제40권1호
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• pp.60-62
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• 1998

To understand expression of some early and late genes of Bombyx mori nuclear polyhedrosis virus (BmNPV) in the B. mori-derived BmN cell line, the transcripts were analyzed by polymerase chain reaction with synthetic primers. After infection, the transcript of early genes, which include p35, IE1 and helicase p143, was immediately detected in the infected cells. In addition, the transcript of late genes, which include p10 and polyhedrin, was also detected in just-infected cells. In conclusion, our results revealed that transcripts of early and late genes of BmNPV are immediately expressed from the cells after infection.

• 한국응용곤충학회지
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• 제35권3호
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• pp.228-231
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• 1996

파밤나방 유충을 이용한 효율적인 SeNPV의 생산을 위하여, 파밤나방유충의 령기와 접종농도에 따른 병원성과 관련하여 바이러스 생체 증식 효율을 조사하였다. 그 결과, 파밤나방 유충을 이용한 SeNPV의 대량생산은 4령 유충에 1.0$\times${TEX}$10^{6}${/TEX} 다각체/ml 농도로 접종하였을 시 약 86.7%의 사충율과 함께 최대 바이러스 생산성을 보여 가장 효율적임을 알 수 있었다.