• Asian-Australasian Journal of Animal Sciences
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• v.16 no.10
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• pp.1535-1539
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• 2003

Four microsatellite DNA loci BM1818, BM1258, BM1443 and BM1905 associated with the somatic cell counts (SCC) in cow milk were analyzed for genetic variation in 240 Beijing Holstein cows. The PCR amplified products of microsatellites DNA were detected by non-denatured polyacrylamide gel electrophoresis. The number of alleles for BM1818, BM1258, BM1443 and BM1905 were 4, 5, 8 and 6 in Beijing Holstein cows, respectively. The allele size ranges for BM1818, BM1258, BM1443 and BM1905 were 274 bp to 286 bp, 92 bp to 106 bp, 154 bp to 170 bp and 187 bp to 201 bp, respectively. The polymorphism information content/effective number of alleles/heterozygosity for BM1818, BM1258, BM1443 and BM1905 were 0.3869/1.7693/0.4348, 0.5923/2.9121/0.6566, 0.7114/3.9012/0.7437 and 0.5921/2.8244/0.6459. These data showed the microsatellite DNA locus BM1443 has the highest variability, followed by BM1258, BM1905 and BM1818. The results of the least squares means analysis showed as follows: the least squares mean of SCC for BM1818 284 bp/284 bp was significantly lower than that for BM1818 286 bp/286 bp (p<0.05). The least squares mean of SCC for BM1258 100 bp/100 bp was significantly lower than that for BM1258 102 bp/102 bp, 106 bp/106 bp, 106 bp/104 bp, 106 bp/102 bp, 106 bp/100 bp, 104 bp/100 bp (p<0.05). The least squares mean of SCC for BM1443 166 bp/160 bp and 166 bp/166 bp was significantly lower than that for BM1443 170 bp/160 bp, 160 bp/157 bp, 165 bp/160 bp (p<0.05). The least squares mean of SCC for BM1905 187 bp/187 bp was significantly lower than that for BM1905 197 bp/195 bp, 193 bp/187 bp (p<0.05).

• International Journal of Industrial Entomology and Biomaterials
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• v.21 no.2
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• pp.157-162
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• 2010

Open reading frame 35 (bm35) of the Bombyx mori nucleopolyhedrovirus (BmNPV) is a special gene whose homologues are only found in some group-I nucleopolyhedroviruses, suggesting that bm35 plays a specific role in the viral life cycle. This paper described the characterization of BmNPV bm35. Computerassisted sequence analysis shows that a putative RING finger motif is observed in the protein, Bm35 encoded by bm35. The coding sequence of bm35 was amplified and subcloned into the vector pET30a(+) and the $(His)_6$-tagged fusion protein His-Bm35 was expressed in the Escherichia coli BL21 (DE3) LysS cells. The bm35 transcript and Bm35 protein were detected in BmNPV-infected BmN cells at 12~48 h post infection (p.i.) by RT-PCR and Western blot analysis using the polyclonal antibody generated by immunizing a rabbit with purified $(His)_6$-tagged Bm35, suggesting that bm35 is synthesized in the late stage of BmNPV infection cycle. Bm35 was not a structural component associated with budded virus (BV) and occlusion derived virus (ODV). These data indicated that bm35 is a functional gene in the BmNPV life cycle.

• BMB Reports
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• v.39 no.6
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• pp.737-742
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• 2006

Open reading frame 60 of Bombyx mori nucleopolyhedrovirus (Bm60) is located between 56,673 and 57,479 bp in the BmNPV genome which encodes 268 amino acid residues with predicted molecular weight of 31.0 kDa. Bm60 and its homologues have been identified in 11 completely sequenced lepidopteran NPVs. The transcript of Bm60 was detected by RT-PCR at 18-72 h post-infection (p.i.), while the corresponding protein could be detected at 24-72 h p.i. in BmNPV-infected BmN cells by Western blot analysis using a polyclonal antibody against Bm60. The expression of Bm60 was inhibited in the presence of Ara-c, an inhibitor of viral DNA synthesis. These results together indicated that Bm60 was a late gene. The size of Bm60 product was found to be a 31 kDa in BmNPV-infected BmN cells, consistent with predicted molecular weight. Immuno-fluoresence analysis showed that the Bm60 product was first detected in the cytoplasm at 24 h p.i and also located in nucleus during later infection. In conclusion, the available data suggest that Bm60 is a functional ORF of BmNPV and encodes a 31kDa protein expressed in the later stage of infection cycle.

• Digital Contents
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• no.10 s.137
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• pp.110-115
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• 2004

지금까지 BM 특허의 인정여부에서부터, 독점 배타적 효력이 있는 BM 특허권을 취득하기 위한 요건들, 그리고 BM 특허권의 권리범위에 대한 문제점까지 알아봤다. BM 특허권을 둘러싸고 존속기간이나 보호 범위에 부정적 의견이 있긴 하지만 일반 기술에 인정되는 특허권과 동일한 효력이 BM 특허에 인정되는 것은 주지의 사실이므로 BM 특허를 기업의 무기로써 어떻게 이용해야 하는지를 고민해야 할것이고 BM 특허가 미래에는 어떠한 모습으로 존재해야 하는가를 논해야 할 것이다. 기업의 전략으로서의 BM 특허와 미래를 논하기에 앞서, 기업의 이익에 큰 몫을 한 특허권 사례를 통해 다시 한번 특허권의 중요성을 강조하고자 한다. 이하는 저작권 보호 솔루션 업체인 인터트러스트(InterTrust)사의 DRM 특허 사례이다.

• Progress in Medical Physics
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• v.25 no.4
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• pp.225-232
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• 2014

We evaluated the effect of scatter on a build-up region based on the measured percent depth dose (PDD) of high-energy photon beams that penetrated a handmade build-up modifier (BM) as a substitute of bolus. BM scatter factors ($S_{BM}$) were calculated based on the PDDs of photon beams that penetrated through the BM. The calculated $S_{BM}$ values were normalized to 1 at the square field side (SFS) of 30 mm without a BM. For the largest SFS (200 mm), the SBM values for a 6-MV beam were 1.331, 1.519, 1.598, 1.641, and 1.657 for the corresponding BM thickness values. For a 10-MV beam, the $S_{BM}$ values were 1.384, 1.662, 1.825, 1.913, and 2.001 for the corresponding BM thickness values. The BM yielded 76% of the bolus efficiency. We expect BM to become useful devices for deep-set patient body parts to which it is difficult to apply a bolus.

• Journal of Sericultural and Entomological Science
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• v.37 no.1
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• pp.52-56
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• 1995

To investigate the effect of hemolymph of silkworm larvae on the multiplication of Bombyx mori nuclear polyhedrosis virus (BmNPV) in BmN-4 cells, BmN-4 cells were infected with BmNPV, which were sequentially Heated with the hemolymph exracted from B. mori larvae. When the culture media TC-100 containing 3% fetal bovine serum was mixed with 10% hemolymph heated at 65$^{\circ}C$ for 30 minutes, the released polyhedra by multiplication of BmNPV in BmN-4 cells were increased more than those of non-treated. However, multiplication of BmNPV in BmN-4 cells treated with non-heated hemolymph was not effective, since non-heated hemolymph was toxic for the cell growth. The result of plaque assay showed that plaque forming units in BmN-4 cells treated with heated hemolymph are significantly increased, suggesting that efficiency of multiplication of BmNPV in BmN-4 cells is due to increase not of cell growth but of infectivity of BmNPV.

• Journal of Sericultural and Entomological Science
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• v.40 no.2
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• pp.105-110
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• 1998

The baculovirus egt gene encodes an ecdysteroid UDP-glucosyltransferase(EGT) which catalyzes the transfer of glucose from UDP-glucose to the insect moltion hormone ecdysteroid resulting in a functionally inactive ecdysteroid. In baculovirus-infected insect larvae, EGT has been shown block molting and pupation. In this study, we compared the development of 4th and 5th instar silkworm, Bombyx mori, larvae injected with either wild-type bombyx mori nucleopolyhedrovirus (BmNPV) or a mutant BmNPV(BmEGTZ) in which the egt gene was disrupted by the insertion of a lacZ gene cassette. Larvae injected with BmEGTZ died roughly 12 h more rapidly compared to indentical larvae infected with BmNPV. In addition, BmEGTZ- infected larvae prematurely stopped feeding and gain less weight compared to BmNPV-infected larvae. In order to investigate why BmEGTZ-infected larvae died more rapidly than BmNPV-infected larvae, the array of hemolymph proteins in BmEGTZ-or BmNPV-infected larvae were analyzed by SDS-PAGE. The hemolymph of BmEGTZ-infected larvae showed virus-specific proteins, including polyhedrin, about 12 h earlier than the hemolymph of BmNPV-infected larvae

• International Journal of Industrial Entomology and Biomaterials
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• v.15 no.1
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• pp.35-38
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• 2007

In this study, ORF 46 of Bombyx mod nucleopolyhedrovirus(Bm46) fused with EGFP was expressed in Bombyx mod cell lines, larvae and pupae by BmNPV Bacmid system. Bm46 and EGFP were cloned into donor plasmid pFastBacHTb, which was transformed to competent DH10B cells containing helper and BmNPV bacmid by site-specific transposition. Recombinant bacmid was used to transfected BmN-4 cells to produce the recombinant baculovirus vBm-Bm46-EGFP. Recombination virus was injected into silkworm larvae and pupae. The expression of the fusion protein was monitored by examining green fluorescence using a fluorescent microscope. Intense fluorescence in cells and silkworm was observed at 4 days post-infection, indicating the Bm46-EGFP fusion gene was expressed successfully.

• 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.

• Journal of Sericultural and Entomological Science
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• v.33 no.1
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• pp.21-26
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• 1991

Bombyx mori nuclear polyhedrosis virus (BmNPV) was successfully multiplied in the nuclear of BmN4 cells cultured with insect Grace's medium. By electron microscopic observation, the virons had a single nucleocapsid in an envelope. Polyhedral protein synthesis of BmNPV in BmN4 cells was detected at 18 hr p.i. and polyhedral protein was a singlepolypeptide with a M.W of 30 kd. At 48 hr p.i. polyhedra formation was observed by inverted mociroscope and electron microscope. Genome analysis of BmNPV by restriction endonucleases was not revealed the difference between virus produced in vivo and that in vitro.