• International Journal of Industrial Entomology and Biomaterials
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• 제16권2호
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• pp.87-92
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• 2008

Bombyx mori nucleopolyhedrovirus (BmNPV) orf47 gene was characterized for the first time. The coding sequence of Bm47 was amplified and subcloned into the prokaryotic expression vector pET-30a(+) in order to produce His-tagged fusion protein in the BL21 (DE3) cells. The His-Bm47 fusion protein was expressed efficiently after induction with IPTG. The purified fusion protein was used to immunize New Zealand white rabbits to prepare polyclonal antibody. As the genome of BmNPV is available in GenBank and the EST database of BmNPV is expanding, identification of novel genes of BmNPV was conceivable by data-mining techniques and bioinformatics tools. Structural bioinformatics approach to analyze the properties of Bm47 encodes protein.

• International Journal of Industrial Entomology and Biomaterials
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• 제21권1호
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• pp.139-143
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• 2010

The objective of our study was the formulation of a local strain of Mamestra brassicae nucleopolyhedrovirus-K1 (MabrNPV-K1) for the development of viral insecticide to control M. brassicae. To formulate MabrNPV-K1, feeding toxicities of various supplements and ultraviolet (UV)-protection were investigated. Optical brightener Tinopal UNPA-GX (Tinopal) as UV protectant and Bentonite had some toxicity themselves to increase the mortality. The protection of polyhedra from UV light radiation was observed only by Tinopal. The MabrNPV-K1 was formulated as a wettable powder form. The mortality of the formulation was higher and rapid than that of the un-formulated. This suggested the possibility of MabrNPV-K1 formulation as an effective biological control agent for M. brassicae.

• International Journal of Industrial Entomology and Biomaterials
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• 제1권1호
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• pp.53-58
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• 2000

A cDNA encoding the luciferase of firefly Luciola lateralis was cloned downstream from the polyhedrin gene promoter of Bombyx mori nucleopolyhedrovirus and expressed in B. mori cells (BmN-4). The coding soquence for luciferase was inserted into pBmKSK2 rectors) which was reconstructed from the polyhedrin-based transfer vector pBmKSKl by modifying cloning sites. Recombinant virus, BmK2-LUCDF, containing the luciferase gene was selected and purified in BmN-4 cells. The emission of luminescence by luciferase was only detected in BmK2-LUCDF-infected cell extracts. This result indicates that the cloned new luciferase gene of firefly L. lateralis can be expressed efficiently in baculovirus expression system and used as a useful reporter gene.

• 한국잠사학회:학술대회논문집
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• 한국잠사학회 2002년도 Join Meetings of Korean Society of Sericultural Science and Japanse Society of Sericultural Science
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• pp.119-119
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• 2002

No Abstract, See Full Text

• Journal of Microbiology and Biotechnology
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• 제11권4호
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• pp.585-591
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• 2001

To compare the p10 promoter activity of Bombyx mori nucleopolyhedrovirus (BmNPV)K1 and K4, recombinant viruses Bm101-LacZ and Bm104-LacZ with a lacZ gene under the control of each p10 promoter were constructed. The $\beta$-galactosidase activity due to Bm101-LacZ was about 5.5- and 1.1-fold higher than that due to Bm104-LacZ and BmK1-LacZ, respectively. expressing ${\beta}$-galactosidase under the control of a polyhedrin promoter. The recombinant virus BmK1-104LacZ with the same genome structure as Bm101-LacZ, except for a p10 promoter region, produced a similar ${\beta}$-galactosidase activity to that due to Bm104-LacZ and 5.5-fold lower than that due to Bm101-LacZ. The virus yield, expression level of polyhedrin, and polyhedra productivity for each recombinant virus was almost similar. These results suggested that the difference in the expression level of ${\beta}$-galactosidase resulted from a difference in the p10 promoter regions, and that an expression vector using the p10 promoter of BmNPV-K1 could be usefully exploited in the mass production of foreign proteins with silkworm larvae by means of oral ingestion.

• BMB Reports
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• 제33권1호
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• pp.63-68
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• 2000

Most baculoviruses have an ecdysteroid UDP-glucosyltransferase (egt) gene, whose product inactivates ecdysteroid within the infected host. Bomhyx mori larvae infected with BmEGTZ, a mutant B. mori nucleopolyhedrovirus (BmNPV) in which the egt gene has been inactivated, die more rapidly compared to larvae infected with wild-type BmNPV. In this study, the profile of hemolymph proteins, and progression of virus infection in BmEGTZ- and BmNPV-infected B. mori larvae, was analyzed by SDS-PAGE and histochemically. These analyses showed that virus-encoded and virus-induced proteins were expressed quicker in BmEGTZ-infected larvae than in BmNPV-infected larvae. This suggests that the decrease in time to death, following BmEGTZ infection, results from the stimulation of virus-specific protein expression. In order to examine the effect of ecdysteroid on virus transmission, the profile of hemolymph proteins, and progression of virus infection, were analyzed following an ecdysteroid injection of BmEGTZ- or BmNPV-infected larvae. In the BmNPV-infected larvae, ecdysteroid treatment had no apparent effect on hemolymph protein expression. This suggests that the injected ecdysteroid was inactivated by the BmNPV-expressed ecdysteroid UDP-glucosyltransferase. An Ecdysteroid injection into BmEGTZ-infected larvae increased the speed of virus-specific protein expression and virus transmission. These results suggest that ecdysteroid stimulates protein expression, which in tum results in the stimulation of virus transmission.

• 한국응용곤충학회지
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• 제38권2호
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• pp.145-149
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• 1999

야생주 핵다각체병 바이러스의 낮은 병원성에 대해 살충제로서의 파밤나방 핵다각체병 바이러스(Spodoptera exigua nucleopolyhedrovirus: SeNPV)의 병원성 향상을 꾀함과 동시에 재조합 바이러스가 다각체 내에 매립됨으로써 살충제로의 적용시 야외에서의 안정성을 확보할 수 있는 p10 유전자의 프로모터를 이용한 새로운 발현벡터를 개발하기 위하여 국내분리주 SeNPV의 p10 유전자 구조를 분석하였다. 그 결과, SeNPV p10 유전자의 프로모터와 구조유전자 부위를 포함한 545염기서열을 결정하여 기존에 보고된 SeNPV p10 유전자(Zuidema et al., 1993)와 비교한 결과 구조유전자 부위에서는 100%의 상동성을 보였으나 5` 및 3` flanking region의 4개의 염기서열에서 차이를 보였다. Southern hybridization에 의하여 SeNPV전체 genomic DNA상에서 p10 유전자는 Sph I 2.4Kb와 Cla I 4.0Kb 단편내에 각각 존재함을 확인하였으며, p10 유전자를 포함하는 이들 단편을 각각 클로닝하여 제한효소 지도를 작성한다.

• 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.

• 한국응용곤충학회지
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• 제42권2호
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• pp.153-157
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• 2003

담배거세미나방 방제를 위한 핵다각체병바이러스의 병원성 검정결과는 다음과 같다. 국화 유묘에서 병원성 검정은 핵다각체병바이러스 1.0$\times$$10^{6.8}$ PIBs/ml에서 노지의 LT$_{50}$ 은 6.2-5.1일이 소요되어 비닐하우스의 6.9-5.4일 보다 짧았으며, 누적 살충율은 노지에서 10일째에 1.0$\times$$10^{7.8}$ PIBs/ml에서 100%의 살충율을 보였으나 비닐하우스에서는 95%였다. 포장의 살충력은 유기합성농약(endosulfan)이 살포 2일 안에 효과가 나타나는 반면 바이러스에 의한 살충은 4일째부터 나타나기 시작하여 14일까지 지속되었다. 노지포장에서는 SINPV 1.0$\times$$10^{8}$ PIBs/ml의 살충율이 94.0%로 대조약제인 endosulfanfsoppm의 91.4%보다 높았으며, 비닐하우스에서는 1.0$\times$$10^{7}$ PIBs/ml이 92.0%로 가장 높았으나 다른처리와 통계적인 유의차가 없었다. 이상의 결과로 보아 노지포장과 비닐하우스에서 1.0$\times$$10^{7}$ PIBs/ml 이상의 농도로 처리하면 담배거세미나방을 방제할 수 있을 것으로 판단되었다.

• 한국응용곤충학회지
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• 제61권3호
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• pp.449-460
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• 2022

광식성 난방제 해충인 파밤나방(Spodoptera exigua)의 친환경적 방제원으로써 이용을 위해 국내에서 분리된 파밤나방 핵다각체병바이러스(S. exigua nucleopolyhedrovirus K1: SeNPV-K1)의 형태 및 전체 유전체 서열을 분석하였다. SeNPV-K1의 다각체(polyhedra)는 0.6-1.8 um 크기의 부정형으로, 기 보고된 SeNPV와 외형적 차이는 보이지 않았다. 전체 유전체의 염기서열을 분석한 결과, 기 보고된 SeNPV와 비교할 때 145 bp 더 많은 135,756 bp로 확인되었으며, G+C 함량은 44% 였고 상동반복영역은 6개로 두 바이러스간에 차이는 없었다. ORF 분석결과, SeNPV-K1은 기 보고된 것과 비교할 때 2개 더 적은 137개를 가지며, SeNPV-K1에만 존재하는 ORF는 4개가 확인되었다. 이들 4개의 ORF는 비필수 유전자로 바이러스의 특성에는 큰 영향을 주지 않을 것으로 여겨졌다. 유전체의 vista 분석 결과, SeNPV-K1과 기 보고된 SeNPV의 전체 염기서열 유사도가 매우 높은 것으로 확인되었다. 국내에서 처음으로 분석한 SeNPV-K1의 전체 유전체는 기 보고된 SeNPV와 유사한 것으로 나타났으나 서로 다른 분리주로 국내 고유자원임을 확인하였다.