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表面展示鸡传染性支气管炎病毒M41株S1蛋白的重组假型杆状病毒的构建

Construction of a Recombinant Baculovirus Surface Displaying S1 Protein of M41 Strain of Infectious Bronchitis Virus
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摘要 以鸡传染性支气管炎病毒M41毒株基因序列为模板,利用PCR方法扩增膜定位信号缺失的S1基因(dS1),将其亚克隆入杆状病毒表面展示质粒pBACsurf-1中,再次将S1及gp64的基因片段克隆到杆状病毒转移质粒pFastBacTMDual,得到重组质粒pFastBac-gp64-dS1.将该重组质粒转化到DH10Bac感受态细胞中,获得重组穿梭载体Bacmid-gp64-dS1,提取重组Bacmid质粒,以PCR验证其正确性.将阳性重组Bacmid质粒利用脂质体转染Sf9昆虫细胞,获得重组杆状病毒BV-dS1.间接免疫荧光试验表明,该重组杆状病毒可以Sf9昆虫细胞膜表达鸡传染性支气管炎病毒S1蛋白. The S1 （Spike） protein gene without membrane localization signal of infectious bronchitis virus （dS1） was amplified from infectious bronchitis virus （M41） by PCR and sub-cloned into pBACsurf-1. The fusion gene containing S1 and gp64 was then inserted into baculovirus transfer vector pFastBacTM Dual to construct the recombinant plasmid pFastBac-gp64-dS1. Then, the plasmid was transformed into Esch- erichia coli DH10Bac competent cells to obtain the recombinant shuttle vector Baemid-gp64-dS1. Extracted Baemid-gp64-dS1 was transfeeted into Sf9 cells to produce the recombinant baculovirus BV-dS1 by using LipofectamineTM 2000. Indirect immunofluorescent assay indicated that the recombinant baculovirus BV-dS1 could express S1 protein on infected Sf9 cell membrane.
作者 陈筱薇 樊惠英 林文耀 程晓亮 叶昱 陈春丽 廖明
机构地区 华南农业大学兽医学院 四川农业大学动物医学院
出处 《华南农业大学学报》 CAS CSCD 北大核心 2012年第3期398-402,共5页 Journal of South China Agricultural University
基金 国家自然科学青年基金(30800826) 广东省博士启动基金(8451064201001131) 农业微生物学重点实验室开发课题(20090010) 华南农业大学校长基金(5500-K08240)
关键词 鸡传染性支气管炎病毒 M41 表面展示 S1蛋白 重组杆状病毒 infectious bronchitis virus M41 surface display S1 protein recombinant baculovirus
分类号 S852.65 [农业科学—基础兽医学]
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参考文献18

  • 1IGNJATOVIC J, GALLI L. Immune responses to structur- al proteins of avian infectinus bronchitis virus [ J ]. Avian Pathol, 1995,24 ( 2 ) : 313-332.
  • 2SEO H S, WANG 1,, SMITH R. et al. The carboxyl-ter,ni- nal 120-residue polypeptide of ilffectious bronchitis virusnucleocapsid induces cytotoxic T lymphocytes and protects chickens from acute infection[J]. J Virol, 1997,71 (10) : 7889-7894.
  • 3CLLISSON E W, PEI Jian-wu, DZIELAWA J, et al. Cyto- toxic T lymphocytes are critical in the control of infectious bronchitis virus in poultry[ J]. Dev Comp Immunol,2000, 24(2/3) :187-200.
  • 4KWON H M, JACKWOOD M W. Molecular cloning and sequence comparison of the S1 glycoprotein of the Gray and JMK strains of avian infectious bronchitis virus [ J ]. Virus Genes, 1995,9 ( 3 ) : 219- 229.
  • 5MARKOVIC I, PULYAEVA H, SOKOLOFF A. Membrane fusion mediated by baculovirus gp64 involves assembly of stable gp64 trimers into muhiprotein aggregates [ J ]. J Cell Biol, 1998,143 ( 5 ) : 1155-1166.
  • 6YANG Ding-gang, CHUNG Yao-chi, LAI Yiu-kay, et al. Avian influenza virus hemagglutinin display on baculovirus envelope: cytoplasmic domain affects virus properties and vaccine potential [ J ]. Mol Ther ,2007,15 ( 5 ) :989-996.
  • 7XU Xin-gang, TANG De-wen, CHIOU Ming-tong, et al. Baculovirus surface display of NS3 nonstructural protein of classical swine fever virus [ J ]. J Virol Methods, 2009,159 (2) :259-264.
  • 8GELB J, Jr, KEELER C L,Jr, NIX W A, et al. Antige- nisl and S-1 genomic characterization of the delaware vari- ant serotype of infectious bronchitis virus [ J ]. Avian Dis, 1997,41 (3) :661-669.
  • 9GELB J,WEISMAN Y,LADMAN B S, et al. S1 gene characteristics and efficacy of vaccination against infectious bronchitis virus field isolates from the United States and Is- rael (1996 to 2000) [J]. Avian Pathol,2005,34(3 ) :194- 203.
  • 10FAN Hui-ying, PAN Yong-fei, FANG Liu-rong, et al. Construction and immunogenicity of recombinant pseudo- type baculovirus expressing the capsid protein of porcinecircovirus type 2 in mice [ J ]. J Virol Methods,2008,150 (1/2) :21-26.

二级参考文献21

  • 1田占成,孙永科,王云峰,智海东,刘胜旺,孙惠玲,王玫,童光志.表达鸡传染性支气管炎病毒S1基因重组鸡痘病毒对SPF鸡的免疫保护作用[J].畜牧兽医学报,2006,37(6):580-586. 被引量：17
  • 2Moennig V,Floegcl Niesmann G,Greiser Wilkc I. Clinical signs and epidemiology of classical swine fever: a review of new knowledge [J].Vet J,2003,165(1):1-2.
  • 3Terpstra C. Hog cholera: an update of present knowledge [J]. Br Vet J,1991,147(5):397-406.
  • 4Van Regenmortel M H V,Fauquet C M,Bishop D H L,et al. Virus taxonomy:the classification and nomenclature of viruses [C]. Report of international committee on taxonomy of viruses. San Diego: Academic Press, 2000.
  • 5Rumenapf T,Unger G,Strauss J H,et al. Processing of the envelope glycoprotdns of pestiviruses [J ]. J Viroh 1993,67 ( 6 ) : 3288-3294.
  • 6Konig M, Lengsfeld T, Pauly T, et al. Classical swine fever virus: independent induction of protective immunity by two structural glycoproteins [J]. J Virol,1995,69(10) :6479-6486.
  • 7Weiland E, Ahl R, Stark R, et al. A second envelope glycoprorein mediates neutralization of a Pestivirus, hog cholera virus [J]. J Virol,1992,66(6) :3677-3682.
  • 8Rumenapf T,Stark R,Mcyers G,et al. Structural proteins of hog cholera virus expressed by vaccinia virus:further characterization and induction of protective immunity [J]. J Virol, 1991,65(2):589-597.
  • 9Van Zijl M,Wensvoort G,de Kluyver E,et al. lave attenuated pseudorabies virus expressing envelope glycoprotein E1 of hog cholera virus protects swine against both pseudorabies and hog cholera [J]. J Virol,1991,65(5) :2761-2765.
  • 10Yang D G,Chung Y C,I.ai Y K,ct al. Avian influenza virus hemagglutinin display on baculovirus envelope: cytoplasmic domain affects virus properties and vaccine potential [J]. Mol Ther.2d07,15(5):989-996.

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华南农业大学学报
2012年 第3期

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