• 제목/요약/키워드: Bracovirus

검색결과 17건 처리시간 0.023초

A SERI technique reveals an immunosuppressive activity of a serine-rich protein encoded in Cotesia plutellae bracovirus

  • Barandoc, Karen P.;Park, Jay-Young;Kim, Yong-Gyun
    • BMB Reports
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    • 제43권4호
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    • pp.279-283
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    • 2010
  • Polydnavirus genome is segmented and dispersed on host wasp chromosome. After replication, the segments form double- stranded circular DNAs and embedded in viral coat proteins. These viral particles are delivered into a parasitized host along with parasitoid eggs. A serine-rich protein (SRP) is predicted in a polydnavirus, Cotesia plutellae bracovirus (CpBV), genome in its segment no. 33 (CpBV-S33), creating CpBV-SRP1. This study explored its expression and physiological function in the diamondback moth, Plutella xylostella, larvae parasitized by C. plutellae. CpBV-SRP1 encodes 122 amino acids with 26 serines and several predicted phosphorylation sites. It is persistently expressed in all tested tissues of parasitized P. xylostella including hemocyte, fat body, and gut. Its physiological function was analyzed by injecting CpBV-S33 and inducing its expression in nonparasitized P. xylostella by a technique called SERI (segment expression and RNA interference). The expression of CpBV-SRP1 significantly impaired the spreading behavior and total cell count of hemocytes of treated larvae. Subsequent RNA interference of CpBV-SRP1 rescued the immunosuppressive response. This study reports the persistent expression of CpBV-SRP1 in a parasitized host and its parasitic role in suppressing the host immune response by altering hemocyte behavior and survival.

A Technique of Segment Expression and RNA Interference (SERI) Reveals a Specific Physiological Function of a Cysteine-Rich Protein Gene Encoded in Cotesia plutellae Bracovirus

  • Barandoc, Karen;Kim, Yong-Gyun
    • Journal of Microbiology and Biotechnology
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    • 제19권6호
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    • pp.610-615
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    • 2009
  • As a provirus, polydnavirus has a segmented DNA genome on chromosome(s) of host wasp. It contains several genes in each segment that presumably play critical roles in regulating physiological processes of target insect parasitized by the wasp. A cysteine-rich protein 1 (CRP1) is present in the polydnavirus Cotesia plutellae bracovirus (CpBV) genome, but its expression and physiological function in Plutella xylostella parasitized by the viral host C. plutellae is not known. This CpBV-CRP1 encoding 189 amino acids with a putative signal peptide (20 residues) was persistently expressed in parasitized P. xylostella with gradual decrease at the late parasitization period. Expression of CpBV-CRP1 was tissue-specific in the fat body/epidermis and hemocyte, but not in the gut. Its physiological function was analyzed by inducing transient expression of a CpBV segment containing CpBV-CRP1 and its promoter, which caused significant reduction in hemocyte -spreading and delayed larval development. When the treated larvae were co-injected with double-stranded RNA of CpBV-CRP1, the expression of CpBV-CRP1 disappeared, whereas other genes encoded in the CpBV segment was expressed. These co-injected larvae significantly recovered the hemocyte-spreading capacity and larval development rate. This study reports that CpBV-CRP1 is expressed in P. xylostella parasitized by C. plutellae and its physiological function is to alter the host immune and developmental processes.

Improved Baculovirus Vectors Expressing Barnase Using Promoters from Cotesia plutellae Bracovirus

  • Choi, Jae Young;Kim, Yang-Su;Wang, Yong;Kang, Joong Nam;Roh, Jong Yul;Shim, Hee Jin;Woo, Soo-Dong;Jin, Byung Rae;Je, Yeon Ho
    • Molecules and Cells
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    • 제28권1호
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    • pp.19-24
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    • 2009
  • The goal of this study was to create a novel baculovirus expression system that does not require recombinant virus purification steps. Transfection of insect cells with transfer vectors containing barnase under control of the Cotesia plutellae bracovirus (CpBV) promoters ORF3004 or ORF3005 reduced cell growth. Co-transfection with bApGOZA DNA yielded no recombinant viruses and nonrecombinant backgrounds. To further investigate the detrimental effects of barnase on insect cells, two recombinant bacmids harboring the barnase gene under control of the CpBV promoters, namely bAcFast-3004ProBarnase and bAcFast-3005ProBarnase, were constructed. While no viral replication was observed when only the recombinant bacmids were transfected, recombinant viruses were generated when the bacmids were co-transfected with the transfer vector, pAcUWPolh, through substitution of the barnase gene with the native polyhedrin gene by homologous recombination. Moreover, no non-recombinant backgrounds were detected from unpurified recombinant stocks using PCR analysis. These results indicate that CpBV promoters can be used to improve baculovirus expression vectors by means of lethal gene expression under the control of these promoters.

프루텔고치벌 브라코바이러스(Cotesia plutellae Bracovirus) 유래 $I_{k}B$ 유전자 구조와 피기생 배추좀나방(Plutella xylostella) 체내 발현 패턴 (Gene Structure of Cotesia plutellae Bracovirus (CpBV)-$I_{k}B$ and Its Expression Pattern in the Parasitized Diamondback Moth, Plutella xylostella)

  • 김용균;;;배성우
    • 한국응용곤충학회지
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    • 제45권1호
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    • pp.15-24
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    • 2006
  • 프루텔고치벌(Cotesia plutellae)은 내부기생봉이고 kB억제자 (IkB)와 유사한 유전자가 이 기생봉의 절대 공생바이러스(C. plutellae bracovirus: CpBV) 게놈에서 발견되었다. 이 유전자의 발현 부위는 417 br의 크기이며 138개 아미노산 서열 정보를 포함하였다. 이 단백질은 4개의 ankyrin 반복영역을 지니고 있었으며, 알려진 다른 폴리드나바이러스 유래 IkB 유전자와 높은 상동성을 보였다. 초파리 Cactus 단백질을 통해 대상 기주 IkB와 비교하여 보면, IkB 신호수신영역이 부재하는 구조를 보여, CpBV-IkB는 NFkB 신호전달체계의 비가역적 억제 인자로 작용할 것으로 추정되었다. CpBV-IkB는 프루텔고치 벌에 기생된 배추좀나방에서만 발현되었다. 정량적 RT-PCR 방법으로 CpBV-IkB의 발현량을 조사하여 보면, 기생 첫날부터 발현을 보이기 시작하여 뚜렷한 발현량을 기생 전체 기간동안 유지하는 양상을 보였다. 이 CpBV-IkB의 기능 분석이 간접적으로 이뤄졌으며, 이 유전자 발현물이 대상 기주 항바이러스 억제 인자로 작용할 것이라는 가설을 제시하였다.

폴리드나바이러스(CpBV) 유래 면역억제 유전자를 이용한 베큘로바이러스 병원력 제고 기술 (Enhanced Pathogenicity of Baculovirus Using Immunosuppressive Genes Derived From Cotesia plutellae Bracovirus)

  • 김용균;권보원;배성우;최재영;제연호
    • 농약과학회지
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    • 제12권3호
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    • pp.283-290
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    • 2008
  • 베큘로바이러스는 일부 나비목 해충을 대상으로 방제하는 데 사용되고 있다. 그러나 화학농약에 비해 느린 살충효과 및 좁은 적용 해충으로 응용 범위에 한계를 갖고 있다. 본 연구는 이러한 한계를 극복하고자 곤충의 면역억제을 통해 바이러스 병원력을 제고시킬 수 있는 기술을 소개한다. 폴리드나바이러스는 일부 맵시벌 및 고치벌에 공생하는 곤충 DNA 바이러스 분류군이다. 프루텔고치벌(Cotesia plutellae) 유래 CpBV(Cotesia plutellae bracovirus)는 브라코바이러스에 속한 폴리드나바이러스로서 면역어제를 발휘하는 여러 유전자를 함유하고 있다. 이 가운데 7개의 CpBV유전자를 선발하고 이를 야생형Autographa California multiple nucleopolyhedrovirus(AcNPV)에 재조합하였다. 이들 재조합 베큘로바이러스를 이용하여 파밤나방(Spodoptera exigua)과 배추좀나방(Plutella xylostella)을 대상으로 생물 검정한 결과, 이들 대부분은 야생형의 바이러스와 유사하거나 우수한 살충력을 나타냈다. 특히 CpBV-ELP를 포함한 재조합 베큘로바이러스가 대조바이러스에 비해 살충시간을 약 2 일 이상단축시킴으로 가장 우수하였다. 이 재조합 베큘로바이러스는 농도에 따른 살충력증가와 배추를 가해하는 파밤나방을 대상으로 한 바이러스 살포 처리가 뚜렷한 방제효과를 나타내어 현장 적용 가능성을 제시하였다. 또한 본 연구는 이 재조합 바이러스의 살충력 제고 현상을 CpBV-ELP의 항바이러스 기작 억제라는 측면에서 고찰했다.