• 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.

• Journal of Microbiology
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• 제41권4호
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• pp.306-311
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• 2003

In Korea, a Lymantria dispar multinucleocapsid nucleopolyhedrovirus, LdMNPV-NM, was isolated and characterized from dead L. dispar larvae. The polyhedra of LdMNPV-NM were irregularly shaped with a diameter of $1.62{\pm}0.33{\mu}m$. Numerous virions comprised of the multinucleocapsid were evident in the electron microscopic examination of the polyhedra cross sections. These polyhedra were composed of a major protein of 30 kDa. The restriction enzyme digestion patterns of LdMNPV-NM showed that this isolate had some different fragments from those of the Gypchek LdMNPV isolate, although their overall profiles were similar. The deduced amino acid sequence of the enhancin gene of LdMNPV-NM showed differences when compared to previously reported enhancin genes of other LdMNPV strains. These results suggested that the LdMNPV-NM isolate from Korea was a new NPV strain and had a new enhancin gene.

• International Journal of Industrial Entomology and Biomaterials
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• 제19권2호
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• pp.237-241
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• 2009

The objective of our study was the evaluation of pathogenicity of a local strain of Mamestra brassicae nucleopolyhedrovirus-K1 (MabrNPV-K1) derived from a diseased larva of M. brassicae found in Korea. The effect of temperature and larval instar on the pathogenicity and production of MabrNPV-K1 was determined under laboratory conditions. The median lethal concentration ($LC_{50}$) values of MabrNPV-K1 for 3rd instar larvae were $3.7\times10^4$ PIBs/larva at $20^{\circ}C$, $9.9\times10^2$ PIBs/larva at $25^{\circ}C$ and $3.8\times10^2$ PIBs/larva at $30^{\circ}C$, respectively. The $LC_{50}$ for the 4th instar larvae was similar to that for the 3rd instar larvae. However, the pathogenicity to the 3rd instar larvae was higher than that to the 4th instar larvae. The median lethal time ($LT_{50}$) values of MabrNPV-K1 were 11.4 to 5.0 days at $30^{\circ}C$ and 18.3 to 5.5 days at $25^{\circ}C$ for the 3rd instar larvae. The $LT_{50}$ value was lowered as temperature went up to $30^{\circ}C$ and dependent on viral concentration. In production efficiency of MabrNPV-K1 using M. brassicae larvae, the mortality of the 3rd instar larvae was 100% when inoculated with $1.0\times10^5$ PIBs/larva and the yield of MabrNPV-K1 was maximal. Regarding the mortality, yield of polyhedra, inoculation doses and required time, the $1.0\times10^4$PIBs/larva at $30^{\circ}C$ was determined as optimal conditions producing polyhedra efficiently.

• International Journal of Industrial Entomology and Biomaterials
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• 제17권1호
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• pp.125-129
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• 2008

The purpose of this study was to investigate the characteristics of Mamestra brassicae nucleopolyhedrovirus (MabrNPV)-K1 isolated in Korea. Polyhedra of MabrNPV-K1 showed irregular appearance in shape with the average diameter $1.8{\mu}m$. MabrNPV-K1 contained a number of nucleocapsids within a viral envelope embedded in polyhedron. The polyhedrin of MabrNPV-K1 was composed of single polypeptide with a M.W. of approximate 31 kDa which is identical to the commercialized MabrNPV, Mamestrin, as a biological control agent. The nucleotide and amino acid sequences within the coding region of MabrNPV-K1 polyhedrin shared 99.0% similarity with the polyhedrin gene from previous reported MabrNPVs. The median lethal concentrations ($LC_{50}$) of MabrNPV-K1 and Mamestrin to M. brassicae larvae were $3.9{\times}10^3$ PIBs/larva and $6.0{\times}10^4$ PIBs/larva, respectively. Mortality of the MabrNPV-K1 against to the third instars larvae was 15 times higher than that of the Mamestrin. The median lethal times ($LT_{50})$ of MabrNPV-K1 by the concentration of polyhedra were lower ($4.4{\sim}6.1$ days) than those of Mamestrin ($4.1{\sim}8.6$ days). These results suggest that a local strain MabrNPV-K1 has high pathogenicity to M. brassicae and may be useful for the development of biological control agent to control this.

• 한국미생물·생명공학회지
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• 제29권1호
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• pp.18-24
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• 2001

누에 핵다각체병 바이러스의 gp64 프로모터를 이용한 transient 발현 벡터를 제작하기 위해서 gp64 유전자의 구조를 분석하였다. Southern blotting 분석을 통해 genome DNA에서 gp64 유전자를 탐색하기 gp64 구조유전자를 포함하는 2,277 nucleotide의 염기를 분석하였으며 gp64의 early, late 프로모터 발현을 조절하는 인자들을 확인하였다. gp64프로모터를 이용한 transient 발현 벡터를 제작하고 외래유전자로써 lacZ 유전자를 Bm5 세포주에서 transient 발현시켰다. 세포주 내에 도입된 플라스미드 DNA의 안정성을 확인하였으며, gp64 프로모터의 외래유전자 발현성 여부를 조사하기 위하여 gp64 프로모터 하에 laxZ 유전자를 가지는 재조합 바이러스를 제작하고 $\beta$-galactosidase in 냐셔 staining을 수행한 결과 전체적인 발현량은 매우 약한 것으로 판단되었다. BmNPV-K1의 gp64 프로모터를 이용한 벡터는 더욱 민감한 표지 유전자를 발현시켜 재조합 바이러스의 분리에 이용하거나 숙주세포에 독성을 보이는 유전자 산물의 소량 발현에 더욱 유용할 것으로 판단된다.

• 한국응용곤충학회지
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• 제43권4호
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• pp.329-332
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• 2004

파밤나방 핵다각체병바이러스(Spodoptera exigua nucleopolyhedrovirus: SeNPV)의 온도별, 농도별로 병원활성을 조사한 결과는 다음과 같다. SeNPV의 온도에 따른 $LC_{50}$은 2령 $20^{\circ}C$에서는 $9.797{\times}10^{3}PIBs/mL$, $32^{\circ}C$에서는 $3.351{\times}10^{2}PIBs/mL$로 온도가 높아질수록 낮아졌으며, 다른 영기에서도 같은 경향이었다. $20^{\circ}C$에서 SeNPV $1.0{\times}10^{3}PIBs/mL$의 파밤나방 3령 유충에 대한 $LT_{50}$은 9.0일이었으나 SeNPV $1.0{\times}10^{4-6}PIBs/mL$에서는 6.9일-3.5일로 더 짧았다. 24, 28, $32^{\circ}C$의 SeNPV $1.0{\times}10^{4}PIBs/mL$에서는 5.7일, 5.5일, 4.9일의 병원 활성을 보여 온도가 올라갈수록 $LT_{50}$은 짧아졌다. 또한 바이러스 농도가 높고 유충의 영기가 어릴수록 짧아져 $LT_{50}$은 온도 바이러스 농도, 유충영기에 따라 차이가 있었다.

• 한국잠사곤충학회지
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• 제50권2호
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• pp.53-62
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• 2012

누에 BmNPV는 잠사업에 있어서 가장 위해한 바이러스로써 익히 보고되었으며, 종종 잠사업의 심각한 경제적 손실을 야기하기도 한다. 곤충의 박테리아, 곰팡이 그리고 원생동물과 같은 다양한 병원체에 대응하는 곤충의 생체 방어기작에 대한 연구가 많이 알려져 있지만, 항바이러스 기작에 대한 연구는 매우 부족한 실정이다. 따라서 본 연구에서는 누에서 처음으로 누에의 BmNPV에 대한 생체방어 관련 유전자를 선발하기 위하여, 누에에 인위적으로 BmNPV를 주사하여 면역을 유도한 다음, 이로부터 cDNA 유전자은행을 제작하였다. 제작된 cDNA 유전자은행으로부터 무작위로 3,332개의 cDNA 클론을 선발하여 정상 누에에 비하여 BmNPV에 의해 면역이 유도된 누에에서 차별화 발현되는 109종의 잠정 항바이러스 유전자 클론을 차별화선별법에 의해서 분리하였다. 본 연구를 통해 확보한 109개의 유전자 정보는 누에의 바이러스에 대한 면역반응뿐만 아니라 최근에 개발된 누에 형질전환 기술을 이용하여 BmNPV 저항성 누에 품종을 개발하는데 중요한 기초 자료를 제공할 것으로 기대되며 또한, 인간의 중요한 항바이러스제 개발을 위한 모델 곤충으로써 누에를 이용하는데 기초 자료로 활용될 것으로도 기대된다.

• 한국응용곤충학회:학술대회논문집
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• 한국응용곤충학회 2001년도 합동 춘계 학술발표회
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• pp.110-110
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• 2001

• 한국응용곤충학회:학술대회논문집
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• 한국응용곤충학회 2004년도 춘계 학술발표회
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• pp.172-172
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• 2004

• 한국잠사학회:학술대회논문집
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• 한국잠사학회 2001년도 제44회 춘계 학술연구 발표자료
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• pp.31-31
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• 2001

No Abstract. See Full-text