• International Journal of Industrial Entomology and Biomaterials
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• 제21권1호
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• pp.117-125
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• 2010

The tropical tasar silkworms, Antheraea mylitta (D) produce famous silk 'Kosa' in central part of India. Due to outdoor rearing it became susceptible to viral infection including cytoplasmic polyhedrosis virus (CPV). The common mode of entry of cytoplasmic polyhedrosis virus is per os and cause gresserie disease to the larvae. Histopathological studies elucidated the insect CPV virus produces infective polyhedral inclusion bodies (PIBs) in the midgut cell cytoplasm of virus infected fifth instar larvae. The PIBs multiply enormously in the cytoplasm without invading the nucleus. Ultrastructural studies confirmed the pathological effects of CPV on in midgut cell cytoplasm. The multiplication of polyhedral inclusion bodies took place into the vacuoles and form virogenic stromata in the cytoplasm of cells. However, the encapsulations of polyhedral inclusion bodies into the polyhedrin protein occurred and polyhedra were released into the lumen. At the late stage of infection, cells showed the regressed cytoplasmic organelles with large vacuoles and elongated mitochondria. Hence, the horizontal transmission of CPV causing the midgut cells disintegration in the tasar silkworm, Antheraea mylitta (D) confirmed during infection.

• Biotechnology and Bioprocess Engineering:BBE
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• 제9권5호
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• pp.352-355
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• 2004

Heliothis zea (HzAM1) insect cells were immobilized in microspheres by sodium-cellulosesulfate (NaCS) and polydiallyldimethylammoniumchloride (PDADMAC). The highest HzAMl cell density was $7.5{\times}10^7$ cells/mL in the microspheres. After infection of the immobilized cells by Heliothis zea single nuclear polyhedrosis virus (HzSNPV), the highest concentration of HzSNPV (polyhedral inclusion bodies: PIBs) produced was $2.87{\times}10^{10}$ PIBs/mL in the microspheres.

• Journal of Microbiology and Biotechnology
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• 제14권2호
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• pp.395-400
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• 2004

Choristoneura fumiferana (Cf-2Cl) insect cells were cultured and immobilized by using cellulosesulfate (NaCS) and polydiallyldimethylammoniumchloride (PDADMAC). A concentration of CfMNPV (Choristoneura fumiferana multiple-mucleopolyhedrovirus) and a Cf-2Cl cell density in the microspheres have been achieved at the densities of $1.57\times10^{10}$ PIBs/ml and $7.5\times10^7$ cells/ml, respectively. Additionally, MTT-test was used to measure the viable cell density in the microspheres, and the confidence of MIT-test was investigated before and after baculovirus infection in the immobilized cell culture.

• 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.

• Journal of Microbiology and Biotechnology
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• 제17권4호
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• pp.632-637
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• 2007

A cytoplasmic polyhedrosis virus (CPV) was isolated from the larvae of Thaumetopoea pityocampa and shown to cause an infection of midgut cells. This viral infection revealed several important diagnostic symptoms, including discoloration of the posterior midgut, reduced feeding, and extended development time of the larvae. The virus infection is lethal to Thaumetopoea pityocampa, and with the increasing doses kills the larvae within 4-5 days post infection. Electron microscopy studies showed typical cytoplasmic polyhedral inclusion bodies that are icosahedral, and ranged from 2.4 to $5.3{\mu}m$ in diameter. Electrophoretic analysis of the RNA genome showed that the virus has a genome composed of 10 equimolar RNA segments with the sizes of 3,907, 3,716, 3,628, 3,249, 2,726, 1,914, 1,815, 1,256, 1,058, and 899 bp, respectively. Based on morphology and nucleic acid analysis, this virus was named Thaumetopoea pityocampa cytoplasmic polyhedrosis virus (TpCPV), and belongs to the genus Cypovirus, family Reoviridae.

• Journal of Microbiology and Biotechnology
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• 제16권10호
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• pp.1485-1490
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• 2006

In this study, comparative replicational properties of Hyphantria cunea nucleopolyhedrovirus (HycuNPV) in Lymantria dispar (IPLB-LdElta) and Spodoptera frugiperda (IPLB-Sf21) cell lines were investigated. Our microscopic observations showed that cytopathic effects (CPEs) in LdElta cells appeared 12 h later than those in Sf21 cells. Whereas polyhedral inclusion bodies (PIBs) formed at 48 h postinfection (p.i.) in LdElta cells, it formed at 36 h p.i. in Sf21 cells. Extracellular virus production determined according to the 50% tissue culture infective dose ($TCID_{50}$) method in LdElta cells started about 12 h later when compared with Sf21 cells. Titers of extracellular virus in LdElta and Sf21 cells were calculated as $1.77{\times}10^9$ plaque forming units (PFU)/ml and $5.6{\times}10^9PFU/ml$, respectively, at 72 h p.i. We also showed that viral DNA replication began at 12 h p.i. in both cell lines. Viral protein synthesis was determined by SDS-polyacrylamide gel electrophoresis (PAGE) and polyhedrin synthesis was observed at 12 h p.i. in both cell lines. The results indicate that while the synthesis of macromolecules is 12 h later and production of extracellular virus is almost 3-fold lower in LdElta cells compared with those in Sf21 cells, the LdElta cell line is still a productive cell line for infection of HycuNPV.

• 한국응용곤충학회지
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• 제42권2호
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• pp.159-163
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• 2003

담배거세미나방 핵다각체병바이러스의 온도별, 농도별로 병원성 검정결과는 다음과 같다. SINPV의 온도에 따른 담배거세미나방 유충 영기별 LC$_{50}$ 은 20-28$^{\circ}C$범위에서는 1,3,5령에서 온도가 높아질수록 낮아졌으나 32$^{\circ}C$에서는 높아졌다. LT$_{50}$ 은 핵다각체병바이러스 1.0$\times$$10^{5.7}$ Polyhedral inclusion bodies (PIBs)/ml에서 온도조건(20-32$^{\circ}C$)이 2$0^{\circ}C$와 24$^{\circ}C$에 비하여 28$^{\circ}C$와 32$^{\circ}C$에서 짧았다. 또한 바이러스 농도가 높고, 유충의 영기가 어릴수록 짧아져 LT$_{50}$ 은 온도조건, 바이러스 농도, 유충 영기에 의존하였다.

• 대한바이러스학회지
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• 제27권2호
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• pp.239-255
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• 1997

전염성 췌장괴저바이러스 (Infectious pancreatic necrosis virus) DRT 주의 VP1유전자를 대 장균 발현운반체와 Baculovirus에 삽입하여 대장균과 진핵세로에서 VP1단백질의 발현을 연구하였다. 재조합체 pMal-pol 클론 [7]에서 2.7 Kb 단편인 VP1 유전자를 제한효소 XbaI으로 절단하여 Baculovirus 운반체인 pBacPAK9에 클로닝하여 pBacVP1이라 명명하였다. 이 pBacVP1에 클로닝된 VP1유전자를 제한효소 SacI과 PstI으로 절단하여 대장균 발현 운반체인 pQE-30에 클로닝하여 pQEVP1이라 명명하였다. 또한 VP1 단백질의 C-말단에 6개의 히스티딘 $6{\times}His$이 붙어 있는 단백질을 만들기 위하여, pQEVP1 클론의 His부위를 EcoRI으로 절단하고, 또한 pBacVP1을 EcoRI으로 절단하여 생긴 부위에His-EcoRI DNA 단편을 교체시켜 재클로닝하여 pBacHis-VP1을 만들었다. pBacHis-VP1 DNA와 Bsu36I로 처리된 LacZ-Hyphantria cunea nuclear polyhedrosis virus (LacZ-HcNPV)를 함께 lipofectin을 이용하여 곤충세포 (Spodoptera frugiperda cell)에 동시 감염을 시켜서 재조합 바이러스를 선발하여, VP1-HcNPV-1이라 명명하였다. pQEVP1 클론은 6개의 히스티딘 단편이 부착된 VP1단백질을 Ni-NTA resin 크로마토그래피법으로 정제하여 SDS-PAGE와 Western blot으로 확인하였고, 단백질의 활성과 구조에 영향을 주지 않는 6개의 히스티딘 단편 ($6\;{\times}\;His$)이 부착된 94 kDa의 VP1단백질을 정제할 수 있었다. 또한 재조합 바이러스에 감염된 곤충세포에서 VP1 단백질이 발현된 것을 전기영동과 Western blot으로 검색을 한 결과 95 kDa VP1 단백질이 발현이 되었음을 확인하였다.