• Journal of Sericultural and Entomological Science
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• v.32 no.2
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• pp.105-109
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• 1990

The disinfectivity of formalin fumigation was tested against silkworm nuclear polyhedrosis virus and yellow muscardine pathogen, Beauveria bassiana. The inactivation of the virus was acquired when it was fumigated by adding 30g of potassium permanganate to 75$m\ell$ of formalin per 3.3$m^2$ of rearing room area and viability of the yellow muscardine pathogen was also lost with the same treatment of fumigation. It was also proved that the fumigation didn't give and damage to silkworm larvae when it was applied 2 or 3 times to grown larvae.

• Korean journal of applied entomology
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• v.36 no.4
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• pp.341-350
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• 1997

We have now constructed a novel recombinant baculovirus producing fusion protein with Autographa californica nuclear polyhedrosis virus (AcNPV) polyhedrin and Bacillus thuringiensis(Bt) cryIA(c) crystal protein. The fusion protein expressed by the recombinant baculovirus in insect cells was characterized. The N-terminal of cryIA(c) gene of Bt subsp. kurstaki HD-73 was introduced under the control of polyhedrin gene promoter of AcNPV, by fusion in the front of intact polyhedrin gene or by insertion into the HindIII site in polyhedrin gene. The recombinant baculoviruses were named as BtrusI or BtrusII, respectively. Although single transcript from the fusion protein gene was apparently observed. BtrusI was produced the two proteins, 92 kDa fusion protein and only polyhedrin. In addition, fusion protein produced by BtrusI did not form polyhedra. Interestingly, however, the cells infected with BtrusII did not show a 33 kDa polyhedrin band as a cells infected with BtrusI. Cells infected with BtrusII were only produced fusion protein, but the polyhedra formed by fusion protein was not observed. To determine the insecticidal toxicity of fusion protein, therefore, Sf9 cells infected with BtrusI were inoculated to Bombyx mori larvae. Sf9 cells infected with BtrusI that expressed the fusion protein caused larval mortality although the insecticidal toxicity was low. In conclusion, our results clearly demonstrated that the fusion protein with polyhedrin and Bt cryIA(c) crystal protein have a insecticida toxicity.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.2
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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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• v.4 no.1
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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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• v.13 no.1
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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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• v.23 no.1
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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.

• Microbiology and Biotechnology Letters
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• v.25 no.4
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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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• v.12 no.1
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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.

• Journal of Sericultural and Entomological Science
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• v.13 no.2
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• pp.141-143
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• 1971

By means of thin-layer electrophoresis in agarose gel, hemolymph protein of healthy silkworm larvae and of the nuclear polygedrosis virus infected larvae were studied. 1. In the 4th instar, 4 fractions moving toward anode were separated. Dye-binding Capacity of the fraction was increased according to the stage. 2. After 5th day in the 5th instar, 7 fractions moving toward anode were separated, and one fraction toward cathode was separated. 3. On the first day in the 5th instar, 5 fractions were separated, and on the 4th day of the same instar 5 fractions were separated. 4. As for the hemolymph protein fractions of the polyhedrosis virus infected larvae, on the 6th and 7th day, three fractions(D.E.F) were inclined to increase, whereas on the 8th day 4 fractions(A.B.D.E) were disappeared but F fraction was inclined to decrease.

• Korean journal of applied entomology
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• v.18 no.1 s.38
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• pp.1-10
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• 1979

An inclusion forming virus isolated from a fan webworm, Hyphantria cunea, in 1975 was identified as a nuclear polyhedrosis virus. With the virus isolated in Korea, it was considered that the virus would be one of the valuable microorganism in microbial control. In this connection, 1) the shape and size of the virus for identification, 2) susceptibility of the various instar larvae to the virus, 3) the effects of storage condition on the pathogenicity and the cross infection of the virus to the larvae of Bombyx mori were examined. The results are summarized as follows; 1. The polyhedron was tetrahedron or hexahedron of $2\mu$ in size and the rod-shaped virus particles consisting of $2\~14$ rods in a bundle were $330m{\mu}\times35m{\mu}$ in size. 2. The hexagonal nuclear polyhedra were found only in the nucleus of the midgut cells but were variable in size. 3. The $LD_{50}$ values for the various instar larvae of H. cunea were $8.377\times10^4\;PIBs/ml$ for the second, $4.974\times10^5\;PIBs/ml$ for the fifth instar larvae. The $LT_{50}values$ for $10^6\;PIBs/ml$ were 9.6 days for the second, 11.5 days for the third, 12.0 days for the fourth and 17 days for the fifth instar larvae. 4. The susceptibility of H. cunea to the nuclear polyhedrosis virus was greater in the first generation than in the second generation. 5. The effect of the storage conditions on the pathogenicity of the nuclear polyhedra was less in refrigerator $(5^{\circ}C)$ and in freezing $(-80^{\circ}C)$ than in room temperature $(18.5^{\circ}C)$, especially as air-dried polyhedra than as suspension. The pathogenicity of the polyhedra seemed to decrease by sunlight during storage as cadavers, since rather greater decrease in pathogenicity was found in sunny condition than in shady condition. 6. The effective spray concentration was $6.4\times10^7\;PIBs/ml$ in the field and its $LT_50$ values for the third and the fifth instar larvae were 4.8 days and 14.2 days, respectively. 7. No cross infections were found in the nuclear polyhedrosis virus between H. cunea and B. mori. larvae.