• Title/Summary/Keyword: Bombyx mori nuclear polyhedrosis virus(BmNPV)

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Biochemical Characteristics of the Nuclear Polyhedrosis Viruses of the Fall Webworm, Hyphantria cunea, and the Silkworm, Bombyx mori (누에와 흰불나방 핵다각체병바이러스의 생화학적 특성)

  • 김현욱;박범석;진병래;임대준;강석권
    • Korean journal of applied entomology
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    • v.28 no.3
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    • pp.105-112
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    • 1989
  • The nuclear polyhedrosis viruses of Bombyx mori (BmNPV) and Hyphantria cunea (HcNPV) were characterized by electron microscopic observation, SDS-PAGE of polyhedral and virion proteins, and restriction endonuclease analysis of viral DNAs. Polyhedra of BmNPV were octadecahedral in shape with the diameter of $3 \mu\textrm{m}$, whereas those of HcNPV showed irregular appearances having the diameter of $1.5~2\mu\textrm{m}$. Under alkaline protease inactivated condition, polyhedral proteins of two NPVs were resolved into a major polypeptide, 30~31 KD, and several minor polypeptides by SDS-PAGE. Examination of virion proteins by silver staining after SDS-PAGE showed that BmNPV was composed of 47 polypeptides with M.W. range of 9.6~112 KD and HcNPV was composed of 48 polypeptides with M.W. range of 9.4~111 KD. The approximate genome size of two NPVs were determined by restriction endonuclease analysis: BmNPV and HcNPV were 114.6 Kb, respectively.

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RNA Interference to Prevent Bombyx mori Nuclear Polyhedrosis Virus Infection in Vivo

  • Hu Zhigang;Chen Keping;Gao Lu;Yao Qin
    • 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.

Location and Nucleotide Sequence of the Bombyx mori Nuclear Polyhedrosis Virus Polyhedrin Gene (누에 핵다각체병 바이러스의 다각체 단백질 유전자의 위치 탐색 및 염기서열)

  • 우수동;김현욱;박범석;강석권;양재명;정인식
    • Journal of Sericultural and Entomological Science
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    • v.34 no.2
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    • pp.20-25
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    • 1992
  • The location of the polyhedrin gene of Bmbyx mori nuclear polyhedrosis virus(BmNPV) was determined by using a cloned polyhedrin gene from the Autographa californica nuclear polyhedrosis virus(AcNPV) as a hybridization probe. The 7.4 Kb PstⅠ fragment DNA of Bm-NPV was cloned to plasmid pUC19 vector. A fragment containing this gene was mapped and sequenced in its entire polyhedrin reading frame. Nucleotide sequences comparison of the polyhedrin of the BmNPV to that of previously reported by Ⅰatrou(1985) revealed that the sequence varied in 10 base, Comparison of the amino acid sequence of the two structured gene revealed that coding sequence varied 74 valine to isoleucine, 76 aspargine to serine and 155 methionine to valine.

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Polyhedral Protein Synthesis and DNA Replication of Bombyx mori, Nuclear Polyhedrosis Virus in a B. mori Cell Line (가잠 배양세포에서 핵다각체병 바이러스의 다각체 단백질 합성과 DNA 복제)

  • 진병래;박범석
    • 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.

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Genomic Recombination of Bombyx mori and Autographa californica Nuclear Polyhedrosis Viruses (누에 및 Autographa californica 핵다각체병 바이러스에 대한 유전자 재조명)

  • 우수동;박범석;박지현;정인식;양재명;강석권
    • 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.

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Molecular Cloning and Sequencing of the Ecdysteroid UDP-Glucosyl-transferase Gene, EGT, from Bombyx mori Nuclear Polyhedrosis Virus K1

  • Park, Hye-Jin;Chung, Eun-Hwa;Lee, Kwang-Sik;Han, Ji-Hee;Lee, Seong-Jin;Sohn, Hung-Dae;Jin, Byung-Rae
    • 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.

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Construction of Stably Transformed Bm5 Cells by Using Autographa californica Nuclear Polyhedrosis Virus IE1 Gene

  • Cho, Eun-Sook;Jin, Byung-Rae;Sohn, Hung-Dae;Chol, Kwang-Ho;Kim, Soung-Ryul;Kang, Seok-Woo;Yun, Eun-Young;Kim, Sang-Hyun;Kim, Keun-Young
    • Journal of Sericultural and Entomological Science
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    • v.40 no.2
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    • pp.111-116
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    • 1998
  • To construct transformed Bm5 cells, Autographa californica nuclear polyhedrosis virus (AcNPV)IE1 gene, an immediate early viral gene was firstly used in this study. AcNPV IE1 gene, which shares on 95.3% uncleotide sequence homology with Bombyx mori nuclear polyhedrosis virus (BmNPV) IE1 gene, was isolated and cloned into pBluescript. Neomycin gene from pKO-neo was inserted under the control of the IE1 promoter to yield pAcIE1-neo. The plasmid pAcIE1-neo was transfected into Bm5 or Sf9 cells, and neomycin-resistant cells were selected in TC100 medium containing 10% fetal bovine serum (FBS) and 1 mg/$m\ell$ G418 for two weeks. Individual clones were picked and each was amplified for further characterization. The genomic DNA from neomycin-resistnt cells was isolated and characterized by PCR using AcNPV IE1 gene-specific primers and by Southern blot analysis using neomycin gene probe. We concluded that AcNPV IE1 gene was functional in B. moridrived Bm5 cells as well as Spodaptera frugiperda-derived Sf9 cells to produce stably-transformed insect cells.

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Influence of Alkaline Protease on Polyhedral Proteins of Nuclear Polyhedrosis Viruses Isolated from Three Lepidopterous Insects (수종 나비목 해다각체병 바이러스의 다각체 단백질 특성과 그에 대한 Alkaline Proteaes의 영향)

  • 박범석;김현욱;진병래;임대중;김석권
    • Korean journal of applied entomology
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    • v.27 no.4
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    • pp.211-218
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    • 1988
  • Polyhedral proteins and the endogenous alkaline protease associated with larval-derived polyhedra of nuclear viruses isolated from Spodoptera litura, Bombyx mori, and Hyphantria cunea were investigated. Polyhedral proteins prepared under alkaline protease heat-inactivated condition were separated as one band with 31Kd in S. litura a H. cunea NpV and 30Kd in B. mori NPV by the SDS-polyacrylamide gel electroptoresis. Whereas polyhedral proteins without heat-inactivation were degraded into smaller polypeptides with a certain pattern in alkaline solution. The results of double-immunodiffusion and western blot analysis with antisera against polyhedral proteins indicated that those three polyhedral proteins had common antigenic determinants and the degradation of polyhedral proteins by alkaline protease could be confirmed.

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Inheritance of Resistance to Nuclear Polyhedrosis Virus in Silkworm, Bombyx mori

  • Sen, Ratna;Ashwath, S.K.;Datta, R.K.
    • International Journal of Industrial Entomology and Biomaterials
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    • v.3 no.2
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    • pp.187-190
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    • 2001
  • Inheritance pattern of resistance to Bombyx mori nuclear polyhedrosis virus (BmNPV) was studied in an Indian silkworm stock TX by single back-cross test method. The resistant parent [TX], susceptible parent [HM], their Fl, F2, and Fl progeny back-crossed to TX [BC(R)] and HM [BC(S)] were inoculated per os with a fixed concentration of BmNPV($0.5{\times}10^{th} PIB/ml$) on the first day of second stadium. The cumulative mortality was recorded until day $10^{\times}$ post-inoculation. The results show that the resistance to BmNPV in TX fellow mono Mendelian inheritance pattern. The resistance dominated over the susceptibility at Fl. At F2, the resistant and susceptible offspring segregated in 3:1 ratio whereas at BC(S), the resistant and susceptible offspring segregated in 1:1 ratio. The response of BC(R) was more or less like the resistant parent TX which confirms the involvement of a major dominant gene conferring resistance to BmNPV in TX. The possible mechanism of inheritance of resistance in TX is discussed.

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NADP-Dependent Malate Dehydrogenase Activity and Associated Biometabolic Changes in Hemolyinph and Fat Body Tissues of Silkworm Bombyx mori L. Following Baculovirus Infection

  • Krishnan, N.;Chaudhuri, A.;Sengupta, A.K.;Chandra, A.K.;Sen, S.K.;Saratchandra, B.
    • International Journal of Industrial Entomology and Biomaterials
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    • v.2 no.2
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    • pp.149-153
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    • 2001
  • The influence of baculovirus Bombyx mori Nuclear Polyhedrosis virus (BmNPV) infection on intermediary metabolic pathways in silkworm Bombyx mori L. was investigated. Studies revealed that NADP-linked malate dehydrogenase activity in hemolymph of infected silkworms at 96 hrs post infection (p.i.) with visible symptoms of infection was enhanced in comparison to healthy larvae of the same age. Also, NADP-dependent MDH activity was significantly lower in fat body cytosol of infected larvae at 96 hrs p.i. when compared to healthy larvae. Similarly, some biometabolic parameters like growth, protein content and cholesterol titer were observed to be influenced by baculovirus infection. While the growth of infected larvae was significantly retardedi protein content was also drastically reduced in both hemolymph and fat body tissues. Cholesterol titers however, was enhanced in infected larvae. The results observed herein point to a significant change in the normal biochemical and biometabolic pathways required for growth and development following BmNPV infection.

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