• Title/Summary/Keyword: bombyx mori nuclear polyhedrosis virus

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Application of Neo-PPS Fumigation to the Disinfection of the Silkworm Larvae, Bombyx mori(L.), for the Control of Silkworm Diseases(I) Effect of Neo-PPS Fumigation on the Virus Diseases (Neo-PPS 훈증에 의한 잠체 소독에 관한 연구(I) -누에 바이러스병에 대한 약효-)

  • 임종성;김근영
    • Journal of Sericultural and Entomological Science
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    • v.18 no.2
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    • pp.79-81
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    • 1976
  • Experiments on the disinfection of the silkworm larvae by the application of Neo-PPS fumigation have been carried out for the control of virus diseases. The results obtained are summarized as follows. The disinfection of 2, 4 and 6 hours'fumigation with Neo-PPS (para-formaldehyde) showed an outstanding effect on the inactivation of the both viruses, nuclear polyhedrosis virus and cytoplasmic polyhedrosis virus, without significance in the 2. 4. and 6 hours' treatment.

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Expression of Escherichia coli ${\beta}$-galactosidase Gene by New Transfer Vector of Baculovirus (새로운 Baculovirus 전이벡터를 이용한 Escherichia coli ${\beta}$-galactosidase 유전자의 발현)

  • Woo, Soo-Dong;Kim, Woo-Jin;Kim, Hye-Seong;Jin, Byung-Rae;Kang, Seok-Kwon
    • Microbiology and Biotechnology Letters
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    • v.24 no.1
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    • pp.72-76
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    • 1996
  • To investigate the expression efficiency of new transfer vector of Bombyx mori nuclear polyhedrosis virus (BmNPV), Escherichia coli lacZ gene was inserted into new transfer vector pBmKSK1, under the control of polyhedrin promoter and expressed in BmN-4 cells and larvae of silkworm, Bombyx mori. The recombinant virus containing lacZ gene was isolated from BmN-4 cells coinfected with transfer vectro pBmKSK1-LacZ and wild type BmNPV genome, and analysed by Southern blotting. The expression of ${\beta}$-galactosidase was characterized by SDS-PAGE, Western blotting and ${\beta}$-galactosidase activity assay. The results showed that the level of expression in silkworm larvae was higher than that of BmN-4 cells.

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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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Effect of Hemolymph of Silkworm Larvae on the Multiplication of Bombyx mori Nuclear Polyhedrosis Virus in BmN-4 Cells (누에 핵다각체병 바이러스의 세포증식에 대한 누에 체액의 영향)

  • 우수동;김우진;진병래;강석권
    • 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.

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Screening of Domestic Silkworm Strains for Efficient Heterologous Protein Expression by Bombyx mori Nuclear Polyhedrosis Virus (BmNPV)

  • Jo, Sun Jung;Choi, Ji-Hyun;Kang, Ju-Il;Lim, Jae-Hwan;Seok, Young Sik;Lee, Jae Man;Kusakabe, Takahiro;Hong, Sun Mee
    • International Journal of Industrial Entomology and Biomaterials
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    • v.29 no.2
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    • pp.185-192
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    • 2014
  • Recombinant proteins can be generated quickly and easily in large amounts and at low-cost in silkworm larvae by using Bombyx mori nuclear polyhedrosis virus (BmNPV). We searched for high-permissive silkworm strains that have high production levels of heterologous proteins and are thus suitable for use as biofactories. In this study, we performed the analysis using a BmNPV vector expressing luciferase as a marker, and we confirmed protein expression by evaluating luciferase activity, determined by western blotting and luciferase ELISA, and confirmed transcription expression by semi- and quantitative real time PCR. For the selection of host silkworm strains, we first chose 52 domestic BmNPV sensitive strains and then identified 10 high-permissive and 5 low-permissive strains. In addition, to determine which hybrid of the high-permissive strains would show heterosis, nine strains derived through three-way crossing were tested for luciferase activity by western blotting, and luciferase ELISA. We found a correlation between luciferase activity and luciferase protein expression, but not transcription. There was no noticeable difference in protein expression levels between Jam313 as the high-permissive control strain and the three-way hybrid strains; however, the three-way cross strains showed lower luciferase activity compared with Jam313. In this study, luciferase protein production in the larvae of 52 domestic silkworm strains was elucidated using BmNPV.

Electron Microscopic Observations on the tissues infected with Nuclear Polyhedosis Virus of Bombyx mori during eary stage (가잠 핵다각체병 바이러스 초기감염에 관한 전자현미경적 관찰)

  • 유강선;강석권
    • Journal of Sericultural and Entomological Science
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    • v.32 no.1
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    • pp.49-57
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    • 1990
  • To investigate the pathway of Nuclear Polyhedrosis Virus(NPV) of Bombyx mori in early stage infection of 2nd instar larva, the larval tissues were observed under electron microscope at interval of 6, 12, 24, and 48 hours after virus inoculation. The results are as follows. 1. The intact and enveloped nucleocapsids released from the polyhedra protein in the gut lumen apparently entered with the microvilli. 2. Virus progenies were observed in columnar cell nuclei 24 hours after inoculation, but polyhedra was not seen. The enveloped virus was observed in some of the intercellular spaces between mid-gut cells. 3. Many enveloped virus particles appeared in the basement membrane. These enveloped virus particles passed the basement membrane and gathered at blood cells in heomocoeal. 4. The NPV muliplicates in nuclei of the blood cells and the tracheal cells normally.

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