• 한국잠사곤충학회지
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• 제32권2호
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• pp.105-109
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• 1990

포르말린에 과망간산카리(KMnO4)를 넣어 화학적 반응에 의하여 발생되는 포름알데히드 가스로 누에고름병 바이러스의 불활화와 흰굳음병균에 대한 소독시험을 실시하여 다음과 같은 결과를 얻었다. 1. 누에고름병 바이러스(Nuclear polyhedrosis virus)의 불활화는 잠실면적 3.3$m^2$당 포르말린 25$m\ell$에 과망간산카리 10g을 혼합하여 하룻방 동안 훈연소독을 실시하였을 때 상당한 수준의 소독효과가 있었고 같은 잠실단위 면적에 포르말린 75$m\ell$와 과망간산카리 30g을 혼합하여 하룻밤 동안 훈연상태를 유지하였을 때는 거의 완전한 불활화가 이루어졌다. 2. 노란굳음병균에 대한 살균효과는 잠실면적 3.3$m^2$당 포르말린 50$m\ell$에 과망간산카리 20g을 반응시켜 훈연소독을 실시하였을 때는 처리받은 균의 자람이 상당한 수준 억제되었고 같은 단위면적에 포르말린 75$m\ell$와 과망간산카리 30g을 혼합시켰을 때는 완전한 살균이 이루어졌다. 3. 포르말린 훈연소독에 의한 누에에 미치는 영향은 4령 1일째와 5령 1일째 2회와 4령 1일째, 5령 1,3일째 각 1회씩 3회를 처리하였을 때(훈연소독은 저녁급상후에 실시하고 다음날 아침 급상때까지 유지시킴) 전견종, 견층중, 견층비율, 유충기간(4-5령) 등에서는 무처리와 차이가 없었으나 3회 처리시에는 실품림새율은 무처리에 비하여 약간 떨어졌다.

• 한국응용곤충학회지
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• 제36권4호
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• pp.341-350
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• 1997

Autographa californica 핵다각체병 바이러스(AcNPV)의 다각체 단백질과 Bacillus thuringiensis(Bt) cryIA(c) 내독소 단백질의 융합단백질을 생산하는 새로운 재조합 바이러스를 제작하고, 곤충세포주(Spodoptera frugiperda 9)에서 발현된 융합단백질의 특성을 분석하였다. Bt kurstaki HD-73의 cryIA(c) 내독소 단백질 유전자의 N-발단 AcNPV의 완전한 다각체 단백질 유전자의 앞쪽에 융합함에 의하여 또는 다닥체 단백질 유전자내의 제한효소 HindII부위에 삽입함에 의하여 다각체 단백질 유전자의 프로모터 조절하에 도입하였다. 이렇게 작성된 재조합 바이러스를 각각 Btrusl 또는 BtrusII라고 명명하였다. BtrusI은 분명히 단일 전사체를 보임에도 92kDa의 융합 단백질과 다각체 단백질의 두 단백질을 생산하였다. 또한 Btrusl에 의해 만들어진 융합 단백질은 다각체를 형성하지 않았다. 한편, BtrusII에 의해 감염된 곤충세포주에서는 33kDa의 다각체 단백질은 보이지 않았고 단지 융합 단백질만 생산하였으나 다각체는 형성하지 않았다. 따라서 Btrusl에 의해 생산된 융합 단백질의 독성을 조사하기 위하여, Btrusl으로 감염된 곤충세포주를 2령 누에(Bombyx mori)에 접종한 결과 융합 단백질에 의한 독성이 관찰되었다. 결론적으로 다각체 단백질과 Bt cryIA(c) 내독소 단백질에 의한 융합 단백질이 독성을 가지고 있음을 확인하였다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권2호
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• pp.146-149
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• 2000

Biologically active porcine Interleukin-2(poIL-2) was produced from in vitro and in viva baculovirus expression system, namely the Autographa californica nuclear polyhedrosis virus (ACNPV)-cell culture system and the Hybrid nucler polthedrosis virus (HyNPV)-sillkworm larva system. The concentration of the recombinant poIL-2(rpoIL-2) in the larvae hemolymph was 1 to 3 mg/mL, which was about 7 to 20 times those of the cell culture systems. The level of this expression efficiency is equal to that with transgenic livestock, secretion products in milk.

• International Journal of Industrial Entomology and Biomaterials
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• 제4권1호
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• pp.51-56
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• 2002

A recombinant transfer vector pBacSCF was constructed by inserting huamn stem cell factor (hSCF) cDNA into plasmid pBacPAK8. BmN cells were co-transfected with modified Bombyx mori, nuclear polyhedrosis virus (BmBacPAK) DNA and the recmbinant transfer vector to construct a recombinant baculovirus containing hSCE gene. DNA dot blotting and RNA dot blotting demonstrated that the hSCE gene was contained in the recombinant virus and transcribed. The recombinant baculovirus was infectious to BmN cells and to silkworm. SDS-PAGE analysis showed a specific band of expressed product in the extract of infected cells and in the heamolymph of infected larvae. Bioactivity of the recombinant hSCE was determined with W-1 cell line and MTT colorimetric method in synergy with interlukin-3 (IL-3). These results revealed that the hSCF gene was over-expressed in cultured cells and lavae of silkworm.

• International Journal of Industrial Entomology and Biomaterials
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• 제13권1호
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• pp.31-35
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• 2006

The ubiquitin conjugating enzyme 2 (E2) is core component of ubiquitin proteasome pathway (UPP) which represents a selective mechanism for intracellular proteolysis in eukaryotic cells. The E2 has been implicated in the intracellular transfer of ubiquitin to target protein. We show here the involvement of E2 in antiviral immune of Bombyx mori to Bombyx mori nuclear polyhedrosis virus (BmNPV). In this study, mRNA fluorescent differential display PCR (FDD-PCR) was performed with BmNPV highly resistant silkworm strain NB and susceptible silkworm strain 306. At 24 h post BmNPV infection, FDD-PCR with the arbitrary primer AP34 showed that one cDNA band was down-regulated in the midgut of resistant strain, but highly expressed in susceptible strain. The deduced amino acid sequence of this cDNA clone share 99% identity with the recently published B. mori ubiquitin conjugating enzyme E2 (Genbank NO: DQ311351). Fluorescent quantitative PCR corroborated down regulation of E2 in resistant strain. We there conclude that BmNPV infection evokes strong response of susceptible strain including activation of UPP. BmNPV may evolve escape mechanisms that manipulate the UPP in order to persist in the infected host. In addition, the identification of down-regulation of E2 in resistant strain, as well as structure data, are essential to understanding how UPP operates in silkworm antiviral immune to BmNPV disease.

• International Journal of Industrial Entomology and Biomaterials
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• 제23권1호
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• pp.129-135
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• 2011

Cathepsins are well-characterized proteases that are ubiquitously expressed in lysosomes. Previous work revealed that $Bombyx$ $mori$ cathepsins B and D are expressed in the fat body and undergo decomposition during larval-pupal metamorphosis. Quantitative RT-PCR was performed to detect cathepsin gene expression at the transcription level when challenged by $B.$ $mori$ nuclear polyhedrosis virus (BmNPV), temperature and hormones (20-hydroxyecdysone (20E) and juvenile hormone analogue (JHA)). mRNAs encoding cathepsins B and D were significantly enhanced after the larvae were infected with BmNPV, and the peak of the induction appeared at 1 day before spinning. This attenuated the inducing effect on cathepsin expression caused by infection. Temperature shock induced cathepsin expression at the later stage of the $5^{th}$ instar, and transcription levels varied with development stage and temperature. Cathepsin B and D mRNA expression in the fat body were significantly induced by JHA at the day before spinning, and with 20E, the expression reached a peak at the last day of the $5^{th}$ instar. Cathepsin B and D mRNA expression exhibited detectable changes post-treatment, without significant differences between or among the hormone concentrations.

• 한국미생물·생명공학회지
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• 제25권4호
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• pp.359-366
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• 1997

The usefulness of host range expanded recombinant viruses for economical viral insecticide and expression vector system has been studied. Host range expanded recombinant viruses, RecS-B6 and RecB-8, constructed by cotransfection of Bombyx mori nuclear polyhedrosis virus (BmNPV) and Autographa californica NPV (AcNPV), and a host range expanded AcNPV recombinant, Ac-BH, constructed by substitution of the 0.6Kb fragment of the BmNPV helicase gene were compared. The restriction enzyme digestion patterns showed that RecS-B6 and RecB-8 had expanded host ranges by genomic recombination and were more similar to genome of AcNPV than that of BmNPV. SDS-PAGE and PCR analysis showed that the polyhedrin gene of RecS-B6 and RecB-8 was derived from BmNPV genomic DNA. The morphology of polyhedra of recombinant viruses showed a slight difference between the two host cells, Sf and BmN cells, indicating that the morphology of polyhedra was influenced by host cells. The bioassay data for insect larvae showed that Ac-BH, compared to wild type viruses, had superior pathogenicity against Bombyx mori larvae but inferior pathogenicity against Spodoptera exigua larvae. Although the pathogenicity was lower than that of wild type viruses in both larvae, RecS-B6 showed the pathogenicity in both larvae. These results suggested that Ac-BH was a less useful economical insecticide than random genomic recombinant virus RecS-B6.

• International Journal of Industrial Entomology and Biomaterials
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• 제12권1호
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• pp.29-34
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• 2006

A breeding programme was initiated during 2001 utilizing two polyvoltine silkworm breeds viz. $BL_{69}$, an evolved breed tolerant to high temperature and MAR, comparatively resistant to Bombyx mori nuclear polyhedrosis virus (BmNPV) with the objective to develop robust polyvoltine breeds and hybrids. The breed $NP_1$ was developed by exposing the fifth instar larvae to high temperature $(36{\pm}1^{\circ}C)$, high Relative Humidity ($85{\pm}5%$ R.H.) and inoculating third instar larvae with BmNPV inoculum. At $F_{12}$, the breed was tested for hybrid forming ability utilizing six bivoltine silkworm breeds viz. $CSR_2,\;CSR_4,\;CSR_{17},\;CSR_{18},\;CSR_{19}\;and\;NB_4D_2$. The hybrid $'NP_1{\times}CSR_{17}'$ exhibited its superiority by recording 97.2% survival, 1.892 g cocoon weight, 0.406 g cocoon shell weight, 21.5% cocoon shell ratio, 16.6% raw silk percentage and 890 m filament length whereas the control $(PM{\times}CSR_2)$ has recorded 90.2% survival, 1.599 g cocoon weight, 0.304 g cocoon shell weight, 18.9% cocoon shell ratio, 13.1 % raw silk percentage and 768 m filament length. Commercial exploitation of the new $polyvoltine{\times}bivoltine$ hybrid in sericulture industry has been discussed.

• 한국잠사곤충학회지
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• 제13권2호
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• pp.141-143
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• 1971

누에의 성장에 따른 단백질분획의 변화 및 핵다각체 이병잠의 체액의 단백질분획의 변화를 agarose gel을 지지체로 한 전기영동에 의하여 조사하였던 바 다음과 같은 결과를 얻었다. 1. 4령기에는 4개의 단백질 분획이 나타났으며 령기가 진전됨에 따라서 염색의 농도가 짙어졌다. 2. 5령 1일에는 5개의 분획이, 5영 4일에는 5개의 분획이 나타났다. 3. 5영 5일 이후의 건강잠의 체액중에는 양극쪽으로 이동하는 7개의 단백질 분획과 음극쪽으로 이동하는 1개의 분획이 분리되었다. 4. 핵다각체 Virus를 첨식시킨 이병잠의 체액단백질 중에서 5령 6일 7일에는 D,E,F 분획이 증가되는 경향이 있고 5령 8일에는 A,B,D,E 분획은 소실되고 F분획은 감소되는 경향이 있다.

• 한국응용곤충학회지
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• 제18권1호
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• pp.1-10
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• 1979

1975년 수원 잠업시험장에서 분리한 흰불나방 핵다각체 바이러스 성상과 령기, 화기별 및 병원체 보존조건에 따른 병원성을 검정하기 위하여 핵다각체 바이러스를 현탁액 및 건조시켜 실온$(18.5^{\circ}C)$, 냉장$(5^{\circ}C)$, 냉동$(-80^{\circ}C)$에 각각 보존, 이병사체는 그대로 양지 음와 지에 보존하였고 야외에서의 병원성, 가잠에 대한 교우감염을 검정한 결과 다음과 같다. 1. 흰불나방 핵다각체는 4면체와 6면체로 크기는 약 $2.0{\mu}$이고 바이러스 입자는 간상형의 $2\~12$본으로 $33m\mu\times35m\mu$이었다. 2. 6각형의 흰불나방 중장핵다각체 바이러스는 중장의 핵에서만 발견되었다. 3. 령기별 $LD_{50.}$ 2,3,4 및 5령에 대해 $8.377\times10^4,\;4.974\times10^5,\;2.621\times10^6$ 및 $9.471\times10^6PIBs/ml$ 였으며 $1.45\times10^6\;PIBs/ml$에 대한 $LT_{50}$은 각각 9.6, 11.5, 12.0 및 17.3일 이었다. 4. 핵다각체 바이러스는 1화기 유충보다 2화기 유충에서 감수성이 높았다. 5. 핵다각체 보존은 둔화건조하여 냉동 및 냉장보존하거나 이병사체를 음지에 보존하는 것이 바이러스 활성도의 감소가 적었다. 6. 야외에서의 효과적인 살포농도는 $6.4\times10^7\;PIBs/ml$이며 이의 $LT_{50}$은 3령은 4.8일, 5령은 14.2일이었다. 7. 흰불나방 핵다각체 바이러스와 가잠의 핵다각체 바이러스는 상호 교우감염을 일으키지 않았다.