• 제목/요약/키워드: baculoviruses

검색결과 34건 처리시간 0.028초

파밤나방 핵다각체병바이러스의 병원성과 곤충간상바이러스의 파밤나방에 대한 교차감염에 관한 연구 (Pathogenicity of Spodoptera exigua Nuclear Polyhedrosis Virus and Cross Infection of Baculoviruses to the Beet Armyworm, S. exigua (Lepidoptera: Noctuidae))

  • 임대준;최귀문;강석권
    • 한국응용곤충학회지
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    • 제30권3호
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    • pp.212-218
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    • 1991
  • 파밤나방 핵다각체병ㅂ이러스의 기주곤충 및 나비목해충에 대한 병원성과 10종 baculovirus의 파밤나방 유충에 대한 교차감염을 조사하여 방제에 이용하고자 시험을 수행하였다. 난에대한 중앙치사농도($LC_{50}$)는 2.855$\times$$10^{5}$ 다각체/ml로 유충보다 높았다. 령별 $LC_{50}$은 3령이 1.422$\times$$10^{4}$ 다각체/ml로 1령보다 1.16배, 5령보다 .11배의 높은 병원성을 나타냈다. 각 령의 중앙치사일수($LT_{50}$)은 1.56$\times$$10^{6}$ 다각체/ml에서 4.26~5.04일이었다. 8종 나비목 해충에 대한 파밤나방 핵다각체병바이러스의 병원성은 기주곤충인 파밤나방에서만 인정되었고 10종의 곤충간상바이러스 중 Autographa californica MNPV, Mamestra brassicae MNPV 및 Trichoplusia ni MNPV 등 3종이 파밤나방 유충에 대해 교차감염을 일으켰다.

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숙주범위가 넓어진 유전자 재조합 핵다각체병 바이러스의 분자생물학적 특성 (Molecular Biological Characterization of Recombinant Baculovirus with an Expanded Host Range)

  • 김우진;우수동
    • 한국잠사곤충학회지
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    • 제38권1호
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    • pp.42-47
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    • 1996
  • AcNPV와 BmNPV를 배양세포주에서 동시감염시켜 선발한 숙주범위가 넓어진 재조합 바이러스인 RecB-727과 RecS-A6의 분자생물학적인 특성들을 조사하였다. 재조합 바이러스의 LT50 값을 조사한 결과, RecS-A6는 모바이러스인 BmNPV 보다 비교적 낮은 병원성을 보았으나 RecB-727은 거의 비슷한 수준의 높은 병원성을 나타내었다. 재조합 바이러스 DNA를 분리하여 모바이러스 DNA와 함께 제한효소 패턴을 비교한 결과 DNA 수준에서 재조합이 일어났음을 확인할 수 있었으며, 일부 유전자의 재조합을 예측할 수 있었다. 또한 p10 유전자에 대한 Southern blot 분석 결과 RecB-727의 p10 유전자는 AcNPV에서 유래되었으며, RecS-A6는 BmNPV의 p10 유전자를 갖고 있는 것으로 추정된다. 재조합 바이러스의 숙주범위 확장에 중요한 역할을 하는 것으로 알려진 DNA helicase 유전자 내의 HindIII-SacI 0.6kb 부위에 대하여 약 250 bp의 염기서열을 조사한 결과, 이 부위의 염기서열은 BmNPV helicase의 염기서열과 동일하였다.

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두 종류의 다각체 단백질 유전자를 발현하는 유전자 재조합 핵다각체병 바이러스의 특성 (Characterization of Recombinant Baculovirus Ewpressing Polyhedrin Gene of Bombyx mori and Autographa californica Nuclear Polyhedrois Virus)

  • 김우진;우수동
    • 한국잠사곤충학회지
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    • 제38권1호
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    • pp.36-41
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    • 1996
  • AcNPV 와 BmNPV의 배양세포주에서의 동시감염에 의해 선발된 재조합 바이러스 RecS-A6는 그 다각체 외부 형태가 모바이러스와 다를뿐만 아니라 배양 세포주에 따라서도 그 형태에 차이가 있었다. 이러한 다각체의 특징적인 형태가 나타나는 요인을 다각체 단백질 유전자를 중심으로 조사한 결과 RecS-A6는 AcNPV 와 BmNPV의 다각체 단백질 유전자를 모두 갖고 있는 것이 확인되었으며, 또한 RecS-A6의 다각체를 단백질 전기영동하여 분석한 결과 RecS-A6의 다각체를 단백질 전기영동하여 분석한 결과 AcNPV와 BmNPV의 다각체 단백질이 모두 다각체 형성에 이용되었음을 확인할 수 있었다.

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Screening of silkworm strains for efficient recombinant protein production by Autographa californica nucleopolyhedrosis virus (AcNPV)

  • Park, Yoon Mi;Kim, Kyung A;Kang, Min Uk;Park, Kwan Ho;Nho, Si Kab
    • International Journal of Industrial Entomology and Biomaterials
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    • 제28권1호
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    • pp.10-18
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    • 2014
  • Baculoviruses base vectors come to be regarded as methods for in vivo gene delivery and transient expression to the silkworm. In the case of silkworm, B. mori, two types of baculoviruses, AcNPV (Autographa californica nuclear polyhedrosis virus) and BmNPV (Bombyx mori nuclear polyhedrosis virus), are potentially applicable as vectors. Recently, AcNPV showed promising results with some silkworm strains despite different host-specificities. We searched for a highly-permissive silkworm strain in the B. mori stocks of Kyungpook National University that could produce high levels of recombinant protein. Seventy strains were screened using the recombinant AcNPV/BmA3-Luc virus. Based on the measured luciferase activity, the strains could be divided into three groups, high-, middle-, and low-permissive strains, according to their relative recombinant protein expression levels. At 48 hours post-injection, the luciferase activity in the high-permissive strains was 500-fold greater than that of the low-permissive strains. At 72 hours post-injection, a significant elevation in luciferase activity was observed in the hemocytes of all strains. Then, based on the above results, the High Permissive Strain (HPS) S10 and the Low Permissive Strain (LPS) S39 were pick up and was carried out Dot blotting, RT-PCR and Real time PCR.

Mass Production of HzSNPV Baculoviruses in Immobilized Heliothis zea (HzAM1) Insect Cell Culture

  • Son Jeong Hwa;Buchholz Rainer;Kim Sung-Koo
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제9권5호
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    • pp.352-355
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    • 2004
  • Heliothis zea (HzAM1) insect cells were immobilized in microspheres by sodium-cellulosesulfate (NaCS) and polydiallyldimethylammoniumchloride (PDADMAC). The highest HzAMl cell density was $7.5{\times}10^7$ cells/mL in the microspheres. After infection of the immobilized cells by Heliothis zea single nuclear polyhedrosis virus (HzSNPV), the highest concentration of HzSNPV (polyhedral inclusion bodies: PIBs) produced was $2.87{\times}10^{10}$ PIBs/mL in the microspheres.

Overexpression, Purification, and Characterization of the Herpes Simplex Virus-1 DNA Polymerase-UL42 Protein Complex

  • Song, Byeong-Doo;Lehman, I. Robert
    • BMB Reports
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    • 제31권6호
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    • pp.585-589
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    • 1998
  • The herpes simplex virus type-1 (HSV-1)-encoded DNA polymerase consists of two subunits, the products of the UL30 and UL42 genes. UL30 and UL42 were coexpressed in Sf9 cells infected with recombinant baculoviruses carrying the two genes. The UL30 and UL42 gene products remained tightly associated throughout the purification, which led to a near homogeneous heterodimer composed of the DNA polymerase and UL42 protein. The DNA polymerase-UL42 protein heterodimer, purified from the recombinant baculovirus-infected Sf9 cells, showed the same high degree of processivity of deoxynucleotide polymerization as the enzyme purified from the HSV-1 infected primate cells. Like the latter, it contained a 3'-5' exonuclease activity that specifically hydrolyzes an incorrectly matched nucleotide at the 3' terminus of a primer, thereby contributing to the fidelity of DNA replication.

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Expression of major piroplasm protein(p33)of Theileria sergenti (Korean isolate) and its immunogenicity in guinea pigs

  • Kang, Seung-Won;Kweon, Chang-Hee;Choi, Eun-Jin;Yoon, Yong-Dhuk
    • Parasites, Hosts and Diseases
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    • 제37권4호
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    • pp.277-283
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    • 1999
  • To investigate the development of a subunit vaccine against theileriosis in cattle, the DNA fragments encoding piroplasm surface protein (p33) of Theileria sergenti of a Korean isolate were expressed in baculoviruses. The expressed p33 was characterized by indirect fluorescent antibody (IFA) and western blotting analysis. The expression of p33 was mainly detected on the surface of infected Sf21 cells by IFA. The immunoblotting analysis revealed the presence of a same molecular weight protein band of p33. The antigenicity of expressed polypeptide was further examined through the inoculation of a guinea pig. The sera of guinea pigs immunized with p33 expressed cell Iysate showed similar fluorescent antibody patterns and reacted with the same molecular weight protein of T. sergenti in immunoblotting analysis, thus indicating that this protein can be a promising candidate for a subunit vaccine in the future.

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Helicoverpa armigera Nucleopolyhedrovirus ORF80 Encodes a Late, Nonstructural Protein

  • Wang, Dun;Zhang, Chuan-Xi
    • BMB Reports
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    • 제40권1호
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    • pp.65-71
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    • 2007
  • The Helicoverpa armigera nucleopolyhedrovirus (HearNPV) ORF80 (ha80) has 765 bp encoding a protein with approximately 254 amino acids and a predicted molecular weight of 30.8 kDa. Homologues of ha80 are found in most baculovirus sequences, including those from lepidopteran NPVs, lepidopteran granuloviruses (GVs), hymenopteran baculoviruses, and one dipteran baculovirus, yet their functions remain unclear. In this study we characterized ha80, and showed that it was transcribed late in infected host cells (HzAM1). The product of ha80 was a 31 kDa protein that was not a structural protein of budded virus (BV) or occlusion-derived virus (ODV) particles. Ha80 was first detected in the cytoplasm of infected HzAM1 cells at 12 h p.i., and was observed in the nucleus at later stages of infection, suggesting that it may be involved in transporting viral proteins into the host cell nucleus or play its roles in the nucleus.