• Title/Summary/Keyword: NPV

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$F(ab)_2$-ELISA for the Detection of Nuclear Polyhedrosis Virus of Silk-worm, Bombyx mori L.

  • Sivaprasad, V.;Nataraju, B.;Baig, M.;Samson, M.V.;Datta, R.K.
    • 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.

Expression of Bombyx mori Nucleopolyhedrovirus ORF4 under the Control of BaculoviruS Ie1 Promoter by a Novel Bac-to-Bac/BmNPV Baculovirus Expression System

  • Su, Wujie;Wu, Yan;Wu, Huiling;Wang, Wenbing
    • International Journal of Industrial Entomology and Biomaterials
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    • v.15 no.2
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    • pp.131-135
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    • 2007
  • Open reading frame 4 of Bombyx mori nucleopolyhedrovirus (BmNPV), designated as Bm4, is a gene whose function is completely unknown. With the recently developed BmNPV bacmid and a modified pFastBac1 whose polyhedrin promoter was replaced with ie1 promoter, a recombinant bacmid expressing Bm4-EGFP fusion protein under the control of ie1 promoter in BmN cells was successfully constructed. The result not only showed that the polyhedrin promoter can be replaced efficiently with other promoters to direct the expression of foreign gene in BmN cells by using Bac-to-Bac/BmNPV baculovirus expression system but also laid the foundation for rescue experiment of Bm4 deletion mutant due to the ability of ie1 promoter to direct gene expression throughout the infection cycle.

A Correlation Study on Nursing Professional Values, Department Satisfaction, Sociality, Self-Esteem among Nursing Students (간호학생의 간호전문직관과 전공만족, 사회성, 자아존중감간의 관계)

  • Yeun, Eun-Ja;Kwon, Young-Mi
    • Journal of Korean Academy of Nursing Administration
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    • v.13 no.3
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    • pp.285-292
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    • 2007
  • Purpose: The purpose of this study was to identify the factors influencing the nursing professional values(NPV) of college nursing students. Methods: A convenience sample of 274 subjects was recruited from the department of nursing at a college. The survey was conducted from May 29 to Jun 20 2006. The data was analyzed using descriptive statistics, Pearson correlation coefficients, and stepwise multiple regression with SPSS/WIN 10.1. Results: The average item score of NPV, satisfaction, socialization, self-esteem were 3.60, 3.00, 2.74, and 2.71. The NPV of nursing students showed a significantly positive correlation to speciality satisfaction(r=.28-.46), socialization(r=.13-.46), and self-esteem(r=.12-.15). The significant factors influencing NPV of nursing students were service, curriculum satisfaction, social perception satisfaction, leadership & popularity, and sociability, which explained about 37.5%. Conclusions: Therefore, on the basis of this study's result, when nursing educational programs are developed to make positive NPV, these factors need to be considered.

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Characterization of a Late Gene, ORF60 from Bombyx mori Nucleopolyhedrovirus

  • Du, Meng-Fang;Yin, Xin-Ming;Guo, Zhong-Jian;Zhu, Liang-Jun
    • 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.

Preliminary Report on the Breeding of Robust and Resistant-NPV and High Quality Silkworm Race 'Shengming No.1' for Summer-autumn Rearing

  • Zhao, Yuan;Chen, Kepin;Yao, Qing;Wu, Yang-Chun;Zhang, Jian;Guo, Xijie
    • International Journal of Industrial Entomology and Biomaterials
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    • v.13 no.2
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    • pp.85-95
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    • 2006
  • Seveval Chinese and Japanese varieties with good characters were used in the breeding. After 5 years (15 generations), a pair of robust and high quality silkworm variety with NPV resistance was bred by means of a combination of crossing and pedigree selection complemented by the selection of NPV resistance. The variety was identified jointly nationwide in 2003 and 2004, and appraised by National Mulberry and Silkworm Appraising Committee. Results are as follows: its cocooning rate is over 93%, shell rate 23-25%, filament length 1200-1300 meters, reelability 75-88%, Length of non-broken cocoon filament 900-1100 meters, raw silk rate 17-19%, neatness 95-97 points, and cocoon crop, cocoon shell weight and raw silk weight per 10000 larvae is higher than those of the control variety by 7-10%, 14-19% and 14-18%, respectively. The variety is not only robust, resistant to high temperature and NPV, easy to rear, uniform in hatching, molting and maturing, but also lays more eggs, and its fecundity is high. It is suitable to rear in the Yangtze River Basin, the Yellow River basin and the Pearl River basin of China.

Molecular Cloning and Characterization of Very Late Expression Factor 1 Gene, vlf-1 from Bombyx mori Nuclear Polyhedrosis Virus Kl

  • Park, Hye-Jin;Lee, Kwang-Sik;Cho, Eun-Sook;Yun, Eun-Young;Kang, Seok-Woo;Kim, Keun-Young;Sohn, Hung-Dae;Jin, Byung-Rae
    • 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.

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Pathogenicity and Polyhedra Morphology of Spilarctia obliqua Nucleopolyhedrosis Virus

  • Kumar, Vineet;Gupta, V.P.;Sarkar, A.;Babu, A.M.;Datta, R.K.
    • International Journal of Industrial Entomology and Biomaterials
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    • v.1 no.2
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    • pp.177-181
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    • 2000
  • Spilarctia obliqua(Wlk.) is a serious pest of mulberry which is naturally affected by its nucleopolyhedrosis virus (SoNPV) in field conditions. The polyhedral occlusion bodies (POB's) were hexahedron under scanning and transmission electron microscope and measured 0.42${\mu}{\textrm}{m}$ to 0.67 ${\mu}{\textrm}{m}$ in diameter. The symptoms of NPV infected S. obliqua larvae resembled with that of other NPVs' infected lepidopterous larvae. The pathogenicity and potentiality of this virus against S. obliqua was tested in the laboratory conditions and the results showed 100% mortality in larvae inoculated with SoNPV at 6.23${\times}10^5$ POBs/ml. Therefore, SoNPV appears to have a high potential as a microbial biocontrol agent against S. obliqua larvae.

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Rapid determination of baculovirus titers an antibody-based assay

  • Kwon, M.S.;Dojimal, T.;Park, Enoch-Y.
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.315-319
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    • 2003
  • A novel method is developed to yield virus titers in 10 h, is easy to .perform using 96-well plates, and applicable to both any Autographa californica nucleopolyhyderovirus (AcNPV) and Bombyx mori nucleopolyhedrovirus (BmNPV)-based recombinant baculovirus. This assay uses an antibody to a DNA-binding protein to detect the infected cells via immune-staining. The titer is determined by counting foci produced due to infection of virus under a fluorescent microscopy. The required incubation period was shortened considerably because infected cells expressed viral antigens at the post infection time of 4 h. Therefore, 10 hours were enough to estimate the virus titer including virus infection time, insect cell culture, and estimation of virus titer.

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Detection of Bombyx mori Nuclear Polyhedrosis Virus(BmNPV) in dust of Silkworm rearing room by PCR (PCR법에 의한 잠실먼지 중 핵다각체병 바이러스의 검출)

  • 남성희;한명세
    • Journal of Sericultural and Entomological Science
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    • v.39 no.1
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    • pp.30-35
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    • 1997
  • A rapid and sensitive detection of BmNPV contamination in silkworm rearing room was carried by Plymerase chain reaction(PCR). Silkworm nuclear polyhedra were dissolved for the extraction of viral DNA within 30 minutes followed by the treatment of alkaline solution. The combination of primers of NP3 and NP2 was superior in PCR to the other 7 primers applied. Each primer was designed with 20 base in size and Newly designed NP3 of sense and the already reported NP2 for antisense were better in reaction than other primers. PCR products appeared 500bp in size. And annealing was confirmed proper at 55$^{\circ}C$ condition. Amplifiable template DNA amount was confirmed at least 100 ng to 0.1 ng and regarded as applicative for the assay of silkworm rearing environmental condition of sericultural farm. In case of the detection of BmNPV from the dust, sensitivity by PCR was as high as 1,000,000 times than that of microscopic observation.

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Biochemical Analysis of Anagrapha falcifera NPV Attachment to Spodoptera frugiperda 21 Cells

  • PARK, JIN O;JAI MYUNG YANG;IN SIK CHUNG
    • Journal of Microbiology and Biotechnology
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    • v.9 no.3
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    • pp.361-364
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    • 1999
  • The binding characteristics of Anagrapha falcifera nuclear polyhedrosis virus (AtNPV) to Spodoptera frugiperda 21 (Sf21) cells were investigated. The cells displayed an affinity of 4.7×10/sup 10/M/sup -1/ with about 3,300 binding sites per cell. The biochemical nature of the AfNPV-binding sites on the cell surface was also partially identified. Our findings suggest that the binding-site moiety has a glycoprotein component, but that the direct involvement of oligosacccharides containing N-acetylglucosamine or sialic acid residues in binding is unlikely, and that AfNPV entry into Sf21 cells may be via receptor-mediated endocytosis.

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