对虾白斑综合症病毒结构蛋白VP28的原核表达和性质研究
RESEARCH ON THE PROKARYOTIC EXPRESSION AND CHARACTERISTICS OF VP28,A STRUCTURE PROTEIN OF PENAEUS MONODON WHITE SPOT SYNDROME VIRUS
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摘要: VP28是对虾白斑综合症病毒(White spot syndrome virus,WSSV)的一个重要的囊膜蛋白.为了便于研究VP28在和宿主细胞相互作用过程中扮演的角色,将VP28基因克隆到一个原核表达系统,对原核表达的VP28的特性进行了研究,并制备了抗VP28的多克隆抗体和单链抗体,对原核表达的和天然病毒的VP28蛋白免疫原性进行了比较.结果表示,原核表达的VP28与天然蛋白具有相似的免疫原性.
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关键词:
- 对虾白斑综合症病毒(WSSV) /
- VP28 /
- 原核表达 /
- 噬菌体抗体
Abstract: White Spot Syndrome Virus(WSSV)is an enveloped DNA virus,which causes 100% cumulative mortality of cultured shrimps in 3 4 days. The virus particles contain five major proteins:VP28、VP26、VP24、VP19 and VP15. The mode of entry and systemic infection of WSSV in shrimp and the functions of these virion proteins are not clear. But more and more attention is paid to the function of VP28,which is likely to play a key role in the initial steps of WSSV infection in shrimps. Towards simplifying to study the role of VP28 in the interaction between WSSV and host cells,the gene of VP28 has been cloned into a prokaryotic expression system,induced and expressed for further researching its characteristics.This prokaryoti expression product could not secrete out of the cell membrane,because it is a membrane protein. Rabbit antiserum against prokaryotic expressed VP28 could recognize both prokaryotic expressed VP28 and native WSSV;Mouse antiserum against native WSSV could recognize both WSSV and prokaryotic expressed VP28. A scFv antibody, designated B7. selected from phage display antibody library against WSSV by panning with prokaryotic expressed VP28,could also recognize both prokaryotic expressed VP28 and native WSSV. These data show that the VP28 produced in a prokaryotic expression system has analogous immunogenic characteristics to its native form,although their protein post modification may differ.-
Keywords:
- White Spot Syndrome Virus(WSSV) /
- VP28 /
- Prokaryotic expression /
- Phage antibody
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[1] Fan X,Zhang S C.New approaches in ocean biotechnology[M].Beijing:ocean publishing Company.1999.30-44.89-100.[ 范晓,张士璀.海洋生物技术新进展.北京:海洋出版社,1999,30-44,89-100][2] Miao H Z ,Tong S L.Multiplication of the shrimp baculovirus HHNBV with primary cell cultures from lymphoid organ of Penaeus chinensis[S].Chinese Journal of Biotechnology 2000,16(2):221-224[苗宏志,童裳亮.利用对虾原代细胞增殖对虾杆状病毒HHNBV的研究.生物工程学报,2000,16(2):221-224][3] Hu C Q.Research on Penaeus Monodon white spot disease and its prevention.Thesises of the second Chinese science meeting on the prevention of shrimp diseases and management of environment [M] Publishing house of ocean university of qingdao 1996,46-51[胡超群.斑节对虾病毒性白斑病及其防治的研究,第二届全国人工对虾疾病综合防治和环境管理学术研讨会论文集.青岛海洋大学出版社,1996,46-51][4] Huang J,Song XL,Yu J,et al.Baeuloviral hypodermal and hematopoietic necrosis-study on the pathogen and pathology of the explosive epidemic disease of shrimp Marine Fisheries Research 1995,16(1):1-10.[黄捷,宋晓玲,于佳,等.杆状病毒的皮下及造血组织坏死-对虾爆发性病的病原和病理学[J].海洋水产研究1995,16(1):1-10][5] van Hulten M C,Marcel W,Stephen D G,et al Identification of two major Virion protein genes of white spot syndrome virus of shrimp.Virology 2002,266:227-236[6] van Hulten M C, Witteveldt J,Marjolein S, et al.White spot syndrome virus envelope protein VP28 is involved in the systemic infection of shrimp.Virology 2001,285:228-233[7] van Hulten M C,Witteveldt J.Peters S,et al.The white spot syndrome virus DNA genome sequence [J] Virology 2001.286:7-22[8] Gao H,Yuan B F, Yuan L, et al.A novel method of screening neutralizing antibody against white spot symdrome virus using primary cultured cells[J].Acta Hydrobiologica Sinica 2002,26(3):259-264[高宏,袁必锋,袁丽,等.一种筛选对虾白斑综合症病毒中和抗体的新方法.水生生物学报2002, 26(3):259-264][9] Yang F,He J, Lin X, et al.The complete genome sequence of the shrimp white spot bacilliform virus.J.Virol.2001,75:11811-11820[10] Dai H,Gao H,Zhao X, et al.Construction and characterization of a novel recombinant single-chain variable fragment antibody against white spot syndrome virus from shrimp.J Immuno.Meth.2003,279:267-275 Fan X,Zhang S C.New approaches in ocean biotechnology[M].Beijing:ocean publishing Company.1999.30-44.89-100.[ 范晓,张士璀.海洋生物技术新进展.北京:海洋出版社,1999,30-44,89-100][2] Miao H Z ,Tong S L.Multiplication of the shrimp baculovirus HHNBV with primary cell cultures from lymphoid organ of Penaeus chinensis[S].Chinese Journal of Biotechnology 2000,16(2):221-224[苗宏志,童裳亮.利用对虾原代细胞增殖对虾杆状病毒HHNBV的研究.生物工程学报,2000,16(2):221-224][3] Hu C Q.Research on Penaeus Monodon white spot disease and its prevention.Thesises of the second Chinese science meeting on the prevention of shrimp diseases and management of environment [M] Publishing house of ocean university of qingdao 1996,46-51[胡超群.斑节对虾病毒性白斑病及其防治的研究,第二届全国人工对虾疾病综合防治和环境管理学术研讨会论文集.青岛海洋大学出版社,1996,46-51][4] Huang J,Song XL,Yu J,et al.Baeuloviral hypodermal and hematopoietic necrosis-study on the pathogen and pathology of the explosive epidemic disease of shrimp Marine Fisheries Research 1995,16(1):1-10.[黄捷,宋晓玲,于佳,等.杆状病毒的皮下及造血组织坏死-对虾爆发性病的病原和病理学[J].海洋水产研究1995,16(1):1-10][5] van Hulten M C,Marcel W,Stephen D G,et al Identification of two major Virion protein genes of white spot syndrome virus of shrimp.Virology 2002,266:227-236[6] van Hulten M C, Witteveldt J,Marjolein S, et al.White spot syndrome virus envelope protein VP28 is involved in the systemic infection of shrimp.Virology 2001,285:228-233[7] van Hulten M C,Witteveldt J.Peters S,et al.The white spot syndrome virus DNA genome sequence [J] Virology 2001.286:7-22[8] Gao H,Yuan B F, Yuan L, et al.A novel method of screening neutralizing antibody against white spot symdrome virus using primary cultured cells[J].Acta Hydrobiologica Sinica 2002,26(3):259-264[高宏,袁必锋,袁丽,等.一种筛选对虾白斑综合症病毒中和抗体的新方法.水生生物学报2002, 26(3):259-264][9] Yang F,He J, Lin X, et al.The complete genome sequence of the shrimp white spot bacilliform virus.J.Virol.2001,75:11811-11820[10] Dai H,Gao H,Zhao X, et al.Construction and characterization of a novel recombinant single-chain variable fragment antibody against white spot syndrome virus from shrimp.J Immuno.Meth.2003,279:267-275
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