• 제목/요약/키워드: polyhedrin

검색결과 61건 처리시간 0.029초

Characterization of Mamestra brassicae Nucleopolyhedrovirus (MabrNPV)-K1 Isolated in Korea

  • Lee, Jae-Kyung;Shin, Tae-Young;Bae, Sung-Min;Choi, Jae-Bang;Oh, Jeong-Mi;Koo, Hyun-Na;Kim, Ju-Il;Kwon, Min;Woo, Soo-Dong
    • International Journal of Industrial Entomology and Biomaterials
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    • 제17권1호
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    • pp.125-129
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    • 2008
  • The purpose of this study was to investigate the characteristics of Mamestra brassicae nucleopolyhedrovirus (MabrNPV)-K1 isolated in Korea. Polyhedra of MabrNPV-K1 showed irregular appearance in shape with the average diameter $1.8{\mu}m$. MabrNPV-K1 contained a number of nucleocapsids within a viral envelope embedded in polyhedron. The polyhedrin of MabrNPV-K1 was composed of single polypeptide with a M.W. of approximate 31 kDa which is identical to the commercialized MabrNPV, Mamestrin, as a biological control agent. The nucleotide and amino acid sequences within the coding region of MabrNPV-K1 polyhedrin shared 99.0% similarity with the polyhedrin gene from previous reported MabrNPVs. The median lethal concentrations ($LC_{50}$) of MabrNPV-K1 and Mamestrin to M. brassicae larvae were $3.9{\times}10^3$ PIBs/larva and $6.0{\times}10^4$ PIBs/larva, respectively. Mortality of the MabrNPV-K1 against to the third instars larvae was 15 times higher than that of the Mamestrin. The median lethal times ($LT_{50})$ of MabrNPV-K1 by the concentration of polyhedra were lower ($4.4{\sim}6.1$ days) than those of Mamestrin ($4.1{\sim}8.6$ days). These results suggest that a local strain MabrNPV-K1 has high pathogenicity to M. brassicae and may be useful for the development of biological control agent to control this.

Production of Monoclonal Antibody about Specific Key Enzyme of Hyoscyamine $6{\beta}-Hydroxylase$ (H6H) in Scopolia parviflora

  • Kang, Young-Min;Jung, Hee-Young;Kang, Seung-Mi;Jin, Byung-Rae;Lee, Sang-Chul;Lee, Byung-Hyun;Choi, Myung-Suk
    • 한국약용작물학회지
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    • 제12권2호
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    • pp.135-140
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    • 2004
  • Total RNAs were isolated from cultured roots of Scopolia parviflora, $poly(A)^+$ RNA was obtained through the mRNA purification, cDNA library of Hnh6h was constructed. Recombinant baculoviruses in Spodoptera frugiperda (Sf) cells were constructed by use of the transfer vector pBacPAK, which has the AcNPV sequence under the polyhedrin promoter. The expression vector carrying Hnh6h gene was transferred to S. parviflora and obtained transgenic hairy root lines. Our results confirmed the over expression of the H6H protein was used by anti-pBacPAK about cDNAs of S. parviflora. This study will served for production of tropane alkaloids by metabolic engineering.

Morphological, Phylogenetic and Biological Characteristics of Ectropis obliqua Single-Nucleocapsid Nucleopolyhedrovirus

  • Ma Xiu-cui;Xu Hai-Jun;Tang Mei-Jun;Xiao Qiang;Hong Jian;Zhang Chuan-Xi
    • Journal of Microbiology
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    • 제44권1호
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    • pp.77-82
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    • 2006
  • The tea looper caterpillar, Ectropis obliqua, is one of the major pests of tea bushes. E. obliqua single-nucleocapsid nucleopolyhedrovirus (EcobSNPV) has been used as a commercial pesticide for biocontrol of this insect. However only limited genetic analysis for this important virus has been done up to now. EcobSNPV was characterized in this study. Electron microscopy analysis of the occlusion body showed polyhedra of 0.7 to $1.7\;{\mu}m$ in diameter containing a single nucleocapsid per envelope of the virion. A 15.5 kb genomic fragment containing EcoRI-L, EcoRI-N and HindIII-F fragments, was sequenced. Analysis of the sequence revealed that the fragment contained eleven potential open reading frames (ORFs): lef-1, egt, 38.7k, rrl, polyhedrin, orfl629, pk-1, hoar and homologues to Spodoptera exigua multicapsid NPV (SeMNPV) ORFs 15, 28, and 29. Gene arrangement and phylogeny analysis suggest that EcobSNPV is closely related to the previously described Group II NPV. Bioassays on lethal concentration $(LC_{50}\;and\;LC_{90})$ and lethal time $(LT_{50}\;and\;LT({90})$ were conducted to test the susceptibility of E. obliqua larvae to the virus.

Expression of Cholera Toxin B Subunit and Assembly as Functional Oligomers in Silkworm

  • Gong, Zhao-Hui;Jin, Hui-Qing;Jin, Yong-Feng;Zhang, Yao-Zhou
    • BMB Reports
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    • 제38권6호
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    • pp.717-724
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    • 2005
  • The nontoxic B subunit of cholera toxin (CTB) can significantly increase the ability of proteins to induce immunological tolerance after oral administration, when it was conjugated to various proteins. Recombinant CTB offers great potential for treatment of autoimmune disease. Here we firstly investigated the feasibility of silkworm baculovirus expression vector system for the cost-effective production of CTB under the control of a strong polyhedrin promoter. Higher expression was achieved via introducing the partial non-coding and coding sequences (ATAAAT and ATGCCGAAT) of polyhedrin to the 5' end of the native CTB gene, with the maximal accumulation being approximately 54.4 mg/L of hemolymph. The silkworm bioreactor produced this protein vaccine as the glycoslated pentameric form, which retained the GM1-ganglioside binding affinity and the native antigenicity of CTB. Further studies revealed that mixing with silkworm-derived CTB increases the tolerogenic potential of insulin. In the nonconjugated form, an insulin : CTB ratio of 100 : 1 was optimal for the prominent reduction in pancreatic islet inflammation. The data presented here demonstrate that the silkworm bioreactor is an ideal production and delivery system for an oral protein vaccine designed to develop immunological tolerance against autoimmune diabetes and CTB functions as an effective mucosal adjuvant for oral tolerance induction.

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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    • 제15권2호
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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.