• Korean journal of applied entomology
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• v.32 no.4
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• pp.407-413
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• 1993

Twelve recombinant viruses with wider host range were plaque purified after coinfectian of Autographa cahjornica and Bombyx mOT! NPVs into Sf9 ar BmN-4 cells. Restriction endonucleases analysis of the recombinant's DNAs showed that the recombinatIOn between AcNPV and BmNPV genomes had occurred more than once. When the recombinam RecB-8, derived from BrnN-4 cells, was observed by electron rntcroscopy, the shape of the polyhedron was a regular tetrahedron, and few virions were occluded into a polyhedron.

• Korean journal of applied entomology
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• v.32 no.1
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• pp.51-60
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• 1993

The polyhedrin gene-containing DNA fragment of Hyphantria cunea nuclear polyhedrosis virus (HcNPV) was localized by southern hybridization with Autographa california CPA EcoRI-I fragment (7.3 kb), Bombyx mori NPV PatI-F fragment (7 kb) and synthetic oligonucleotide(30-mer) as probes. the PstI-L(5.3 kb) fragment of HcNPV was cloned to E. coli and the plasmid of the fragment was named as pHcP-L(8.0 kb). The pHcP-L was physically mapped and subcloned to E. coli as pHcP-L1(4.7 kb), pHcP-L2(7.1 kb), pHcP-L3(5.3 kb), pHcP-L4(4.2 kb) and pHcP-L5(4.5 kb).

• International Journal of Industrial Entomology and Biomaterials
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• v.3 no.1
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• pp.37-41
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• 2001

The ecdysteroid UDP-glucosyltransferase (egt) gene isolated from Bombyx mori nuclear polyhedrosis virus (BmNPV) K1 strain was compared to its homologue from Autographa californica NPV (AcNPV) and Bm NPV T3. The egt gene of BmNPV-K1 encoded 506 amino acid open reading frame, and was 99.6% identical at the amino acid level and 99.2% identical at the nucleotide level to BmNPV T3. The BmNPV-K1 egt gene showed highly identity to AcNPV and BmNPV T3 strain. The BmNPV-K1 egt gene was different from amino acid sequence at 2 positions, 19 and 72, in BmNPV T3. The genomic location of egt gene in the BmNPV-K1 was confirmed by Southern blot analysis and its expression patterns at the transcriptional level in the infected cells were confirmed by Northern hybridization analysis. Transcripts of the egt of Bm NPV-K1 peaked around 12 hrs postinfection (p.i.) and reduced at 24 hrs p.i.

• Proceedings of the Korean Society of Sericultural Science Conference
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• 2000.10a
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• pp.100-100
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• 2000

No Abstract.See Full-text

• Journal of Sericultural and Entomological Science
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• v.37 no.1
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• pp.46-51
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• 1995

In order to develope baculovirus expression vector system, we constructed new transfer vector of nuclear polyhedrosis virus of the silkworm, Bombyx mori. The promoter region containing only adenine of translation start codon of polyhedrin gene was cloned by polymerase chain reaction technique. And the 5' and 3' leader regions of polyhedrin gene was sequentially cloned. The polyhedrin coding gene was deleted from the ＋2 to the ＋597 position. As the result, we constructed new transfer vector which has EcoRI, SacI and KpnI sites for the cloning sites of foreign gene. New transfer vector was named as pBmKSKl. Escherichia coli $\beta$-galactosidase gene as foreign gene was inserted into pBmKSKl, under the control of the polyhedrin promoter and expressed in B. mori cells. The result showed that the new transfer vector pBmKSK1 is functional.

• Korean journal of applied entomology
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• v.30 no.2
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• pp.144-149
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• 1991

Biochemical characteristics of Spodoptera uigua nuclear polyhedrosis virus (SeNPV) isolated in Jinju were studied. SeNPV contained a number of nucleocapsids within a viral envelope embeded in polyhedra. The polyhedral protein of SeNPV was composed of a single polypeptide with a M. W. of 30kd. Double-immunodiffusion test showed that the polyhedral protein of SeNPV had common antigenic determinants with SINPV and BmNPV. Virion proteins of SeNPV were resolved into 49 polypeptides by silver staining after SDS-PAGE. The approximate genome size of SeNPV by restriction endonuclease analysis was 1l0kb.

• Journal of Sericultural and Entomological Science
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• v.39 no.2
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• pp.180-185
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• 1997

To develope the novel baculovirus transfer vector, the p10 gene was cloned from the Bombyx mori nuclear polygedrosis virus (BmNPV) vB2 strain isolated from the B. mori larvae of sericultural farms. The novel transfer vector was constructed by using the p10 gene of BmNPV vB2 strain was 210 bp. The TAAG sequence at the -71 bp of upstream from translation initiator ATG and two polyadenylation signal site at the downstream from terminator TAA were also detected in the p10 gene. The 5' and 3' flanking region of the p10 gene amplified by PCR was cloned into pBluescriptII SK(+) and then transfer vector pBm10 was construceted. The 7.9 kb pBm10 was analysed by restriction enzymes and the map was confirmed. In order to determine the expression of foreign gene of pBm10, $\beta$-galactosidase gene was inserted in the SmaI site of foreign gene cloning site of pBm10. The pBm10 containing $\beta$-galactosidase gene was cotranfected wth genomic DNA of BmNPV vB2 into BmN-4 cells. The recombinant baculovirus expressing $\beta$-galactosidase was also produced polygedra in the infected cells. The results indicated that pBm10 is functional, suggesting that in the baculovirus expression vector system, the recombinant virus produced by pBm10 was effective by oral infection for the producing recombinant proteins in in vivo expression.

• International Journal of Industrial Entomology and Biomaterials
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• v.12 no.2
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• pp.95-100
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• 2006

Central Sericultural Research and Training Institute, Mysore have evolved several highly productive bivoltine hybrids which can produce international grade raw silk. Among them $CSR2{\times}CSR4,\;CSR2{\times}CSR5,\;CSR3{\times}CSR6,\;CSR17{\times}CSR16,\;CSR18{\times}CSR19$ and $CSR12{\times}CSR6$ are being popularized in the field. There is a minimum difference in their economic characters but they appear to differ in survival. Though they are productive under high input management conditions, they are very susceptible to different diseases under normal rearing practices. No systematic attempts have been made to test their susceptibility status / resistance. Thus the present study is a modest attempt to screen the above six productive bivoltine hybrids to two important pathogens viz., Bombyx mori Nuclear Polyhedrosis Virus (BmNPV) and Bombyx mori Infectious Flacherie Virus (BmIFV) along with existing hybrid, $KA{\times}NB4D2$ to assess their susceptibility / resistance. The results shows that the productive hybrid $CSR2{\times}CSR4$ is the most resistant to BmNPV and it is suggested by its highest $LC_{50}$ value followed by $CSR12{\times}CSR6,\;KA{\times}NB4D2,\;CSR3{\times}CSR6,\;CSR17{\times}CSR16,\;CSR18{\times}CSR19,\;CSR2{\times}CSR5$. Based on the $LC_{50}$ value and $LT_{50}$ values for BmIFV, the hybrid $KA{\times}NB4D2$ was found to be the most resistant (1st position) one followed by $CSR3{\times}CSR6$ (2nd position) $CSR2{\times}CSR$ (3rd position) and $CSR12{\times}CSR6$ (4th position) $CSR17{\times}CSR16$, $CSR18{\times}CSR19$ (5th position) and $CSR2{\times}CSR5$ being the least. The response of 7 bivoltine hybrids to both the pathogens BmNPV and BmIFV indicates that, the hybrids $CSR2{\times}CSR4$, $CSR12{\times}CSR6$ and $KA{\times}NB4D2$ were found to be the most resistant when compared to others. Further, $KA{\times}NB4D2$ being less productive hybrid with a shell ratio of 20.08%, the other two hybrids $CSR2{\times}CSR4$ (Cocoon shell ratio, 21.44%) and $CSR12{\times}CSR6$ (cocoon shell ratio, 23.45%) can be considered to be most productive with superior quality cocoon and resistant to both BmNPV and BmIFV pathogens. The overall study indicated that the hybrid $CSR2{\times}CSR5$ is the most susceptible hybrid to both the pathogens.

• International Journal of Industrial Entomology and Biomaterials
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• v.1 no.2
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• pp.111-114
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• 2000

The female pupae of the silkworms Bombyx mori, were injected with recombinant Autographa californica nuclear polyhedrosis virus (AcNPV) expressing green fluorescent protein (GFP) by percutaneous inoculation. When the 4 day-old female pupae were injected with 1x10$^{7}$ or 2${\times}$10$^{7}$ plaque forming units (pfu) of the recombinant AcNPV, oviposited number and egg weight were significantly decreased. Furthermore, the shape of the eggs was obviously divides into normal and abnormal shapes. The percentage of the eggs with an abnormal shape was 7.8% and 57.1% at 1${\times}$10$^{7}$ and 2${\times}$10$^{7}$ pfu inoculation, respectively. PCR analysis of the genomic DNA extracted from the eggs revealed that gfp and AcNPV ecdysteroid UDP-glucosyltransferase genes were amplified from both types of eggs with normal and abnormal shapes. The results demonstrate that AcNPV DNA, and gfp gene cloned into the AcNPV genome, injected in pupal stage were transmitted to eggs and remained stable through at least next generation.

• International Journal of Industrial Entomology and Biomaterials
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• v.1 no.2
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• pp.149-153
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• 2000

We have constructed a recombinant baculovirus, Autographa californica nuclear polyhedrosis virus (AcNPV), containing green fluorescent protein (GFP) gene from the jellyfish, Aequorea victoria, and transferred it into the domestic silkworm Bombyx mori larvae for the production of visible transgenic silkworm of living organism. When one day-old fifth instar female larvae were injected with the recombinant AcNPV of 1x10$^{5}$ plaque forming units, the bright glow of GFP was detected in the recombinant AcNPV-infected larvae and in the newly hatched larvae of the next generation. Our findings demonstrate that the viral replication was detected in the silkworm treated with the recombinant ACNPV and the gfp gene was expressed under the transcriptional control of the polyhedrin gene promoter, Furthermore, the gfp gene was transmitted to the next generation, suggesting that this system can be applied for the development of transgenic silkworms.