• International Journal of Industrial Entomology and Biomaterials
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• v.6 no.2
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• pp.179-181
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• 2003

$F(ab`)_2$-ELISA and direct antigen coating-ELISA (DAC-ELISA) were evaluated in the detection of purified Bombyx mori nuclear polyhedrosis virus (BmNPV) and nuclear polyhedrosis virus infection in silkworm larvae inoculated with BmNPV polyhedra. Although nanogram levels of BmNPV was detected in both DAC- and $F(ab`)_2$-ELISA, similar concentrations of antigen was detected in case of F(ab’)$_2$-ELISA even at higher dilution of antibody (up to 1 : 20 K). One hundred percent nuclear polyhedrosis infection was detected 6 hrs after inoculation in BmNPV infected silkworm larvae by $F(ab`)_2$-ELISA. On the other hand, detection of 100% infection was observed only three days after inoculation in DAC-ELISA. In this study, it was observed $F(ab`)_2$-ELISA was more sensitive than DAC-ELISA in the detection of purified BmNPV as well as nuclear polyhedrosis infection in silkworm larvae.

• Journal of Sericultural and Entomological Science
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• v.40 no.1
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• pp.60-62
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• 1998

To understand expression of some early and late genes of Bombyx mori nuclear polyhedrosis virus (BmNPV) in the B. mori-derived BmN cell line, the transcripts were analyzed by polymerase chain reaction with synthetic primers. After infection, the transcript of early genes, which include p35, IE1 and helicase p143, was immediately detected in the infected cells. In addition, the transcript of late genes, which include p10 and polyhedrin, was also detected in just-infected cells. In conclusion, our results revealed that transcripts of early and late genes of BmNPV are immediately expressed from the cells after infection.

• International Journal of Industrial Entomology and Biomaterials
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• v.12 no.1
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• pp.15-19
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• 2006

RNA interference has been used as a powerful tool in preventing virus proliferation in many species. In this study, we injected the dsRNA in vitro transcripts into Bombyx mori to investigate the resistance to B. mori nuclear polyhedrosis virus (BmNPV). Through vivisectional observation and real-time quantities PCR analysis, we found that these dsRNA can prevent the BmNPV to a certain extent, and delay the viruses' proliferation.

• International Journal of Industrial Entomology and Biomaterials
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• v.2 no.2
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• pp.119-122
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• 2001

Whether Bombyx mori nuclear polyhedrosis virus (BmNPV) can be transmitted to offspring, has been a noticeable question for a long time. When fifth instar larvae of the silkworm were orally inoculated with BmNPV dot hybridization and PCR amplification analysis demonstrated that BmNPV was not detected in the eggs laid by BmNPV productively infected female moths. The results indicated that BmNPV could not be transovarially transmitted.

• International Journal of Industrial Entomology and Biomaterials
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• v.1 no.1
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• pp.29-33
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• 2000

We cloned and characterized a very late expression factor 1 gene, vlf-1, which regulates the level of very late gene transcripts, from Bombyx mori nuclear polyhedrosis virus (BmNPV) K1 strain. The 1,140 bp vlf-1 has an open reading frame of 379 amino acid and a MW of 44 kDa. The vlf-1 nucleotide sequence of BmNPV-Kl showed high homology with Autographa californica nuclear polyhedrosis virus and BmNPV T3 strain so far known, and its deduced amino acid residues were identical to those of BmNPV T3. The location of vlf-1 in the BmNPV-Kl genome was confirmed by Southern blot analysis and its expression patterns at the transcriptional level were confirmed by Northern hybridization analysis.

• International Journal of Industrial Entomology and Biomaterials
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• v.7 no.1
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• pp.87-91
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• 2003

BmNPV (Bombyx mori nuclear polyhedrosis virus) causes nuclear polyhedrosis in silkworms. The tolerance of silkworms to BmNPV is controlled by polygenes. This paper reports on the relative tolerance of silkworm breeds among the germplasm maintained at Andhra Pradesh State Sericultural Research ＆ Development Institute (APSSRDI), Hindupur, India. The silkworm larvae out of second moult were per orally inoculated with BmNPV polyhedra $(l{\times}l0^{th}//ml)$ and reared upto spinning. The response to BmNPV had been categorized into apparent tolerance, real tolerance and susceptibility. Among the 145 silkworm breeds screened, 18 bivoltines and 16 polyvoltines were found to have real tolerance to BmNPV.

• International Journal of Industrial Entomology and Biomaterials
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• v.7 no.1
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• pp.5-10
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• 2003

The nuclear polyhedrosis virus (NPV) resistance of silkworm is controlled by a pair of dominant genes on autosome and micro-effect modificator genes on sex chromosome Z and has the phenomenon of patroclinal inheritance. Based on its hereditary characteristics, methods of preparing near isogenic lines and their $F_2$ populations for screening molecular markers were designed.

• International Journal of Industrial Entomology and Biomaterials
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• v.3 no.1
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• pp.43-49
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• 2001

We have cloned and characterized an immediate early-1 gene, iel, which is activated immediately upon entrance of the viral genome into the cell nucleus, from Bombyx mori nuclear polyhedrosis virus (BmNPV) K1 strain. This gene encodes a protein 584 amino acids with a predicted molecular weight of 67 kDa. The promoter and coding regions of BmNPV-K1 ie1 showed high homology with Autographa californica nuclear polyhedrosis virus and BmNPV T3 strain. The BmNPV-K1 ie1 was different from amino acid sequence at 4 positions in BmNPV T3. The location of ie1 gene in the BmNPV-K1 genome was confirmed by Southern blot analysis and its expression patterns at the transcriptional level in the infected cells were confirmed by Nerthern hybridization analysis.

• International Journal of Industrial Entomology and Biomaterials
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• v.3 no.1
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• pp.25-29
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• 2001

We have cloned and characterized an antiapoptotic gene, p35, which blocks apoptosis, from Bombyx mori nuclear polyhedrosis virus (BmNPV) K1 strain. The 897 bp p35 has an open reading frame of 299 amino acids. The BmNPV-K1 p35 showed a high identity to Autographa californica nuclear polyhedrosis virus and BmNPV T3 strain. The BmNPV-K1 p35 was different from the amino acid sequences of BmNPV T3 at 6 positions. The p35 gene of BmNPV-K1 was 99.2% identical at the nucleotide level and 98% identical at the amino acid level to BmNPV T3. The location of p35 gene in the BmNPV-K1 genome was confirmed by Southern blot analysis and its expression patterns at the transcriptional level in the infected cells were con- firmed by Northern hybridization analysis.

• International Journal of Industrial Entomology and Biomaterials
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• v.3 no.1
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• pp.75-81
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• 2001

We have constructed a novel recombinant Bombyx mori nuclear polyhedrosis virus (BmNPV) producing the green fluorescent polyhedra. For the production of the fluorescent polyhedra, partial polyhedrin gene containing KRKK as nuclear localization site from the BmNPV polyhedrin gene and the green fluorescent protein (gfp) gene were introduced under the control of p10 promoter of BmNPV. The recombinant BmNPV was stably produced fluorescent polyhedra in the infected Bm5 cells and the morphology of the fluorescent polyhedra was similar to that of wild-type BmNPV. The fluorescent polyhedra had 32 kDa native polyhedrin and 41 kDa fusion protein. From these data, we have further developed a novel BmNPV p10-based transfer vector producing recombinant polyhedra with foreign gene Product. The novel BmNPV P10-based transfer vector is composed of partial polyhedrin gene, factor Xa, and multiple cloning sites.