期刊文献+
任意字段
题名或关键词
题名
关键词
文摘
作者
第一作者
机构
刊名
分类号
参考文献
作者简介
基金资助
栏目信息

抗蓝舌病病毒VP7和NS2蛋白单克隆抗体的制备及鉴定 被引量：4

Production and identification of monoclonal antibodies against VP7 and NS2 of bluetongue virus
在线阅读 下载PDF
导出
摘要 为建立蓝舌病病毒(BTV)的检测方法和研究该病毒蛋白的功能,本研究利用BTV血清8型(BTV8)免疫BALB/c小鼠,取免疫后的小鼠脾淋巴细胞与SP2/0细胞融合,制备单克隆抗体(MAb)。并以BTV8作为包被抗原建立间接ELISA方法,经筛选获得了8株稳定分泌抗BTV8 MAb的杂交瘤细胞株(1B2、1F6、2B1、2D10、3B6、3D9、4D4和4D12)。Western blot结果显示,MAb 1F6、2B1、2D10、3B6、3D9与BTV8 VP7蛋白反应,MAb B2、4D4、4D12与BTV8 NS2蛋白反应。间接免疫荧光结果显示,该8株MAb与24型BTV血清型呈不同的反应谱系。本研究所获得的MAb为建立BTV免疫学检测方法和相关病毒蛋白的功能研究提供了实验依据。 To prepare monoclone antibody （MAb） against bluetongue virus （BTV）, SP2/0 myeloma cells were fused with spleen cells of BALB/c mice immunized with BTV serotype 8, and screened by indirect ELISA coated with BTV8. Eight MAbs （1B2, 1F6, 2B1, 2D10, 3B6, 3D9, 4D4 and 4D12） against BTV were produced. The result of western blot indicated that the MAbs of 1F6, 2B1, 2D10, 3B6, 3D9 reacted to VP7 protein, and the rest of MAbs reacted to NS2 protein. The results of IFA showed that all the MAbs had different reactivity with BTV1-24 serotypes. The MAbs produced in this study are useful tools for developing immunological diagnostic techniques of BTV and investigating proteins related to BTV.
作者 秦永丽 孙恩成 耿宏伟 杨涛 刘霓红 赵晶 王凌凤 徐青元 蔡绪禹 李文京 吴东来
机构地区 中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/农业部兽医公共卫生重点开放实验室 东北农业大学动物医学学院 中国动物疫病预防控制中心
出处 《中国预防兽医学报》 CAS CSCD 北大核心 2011年第10期816-819,共4页 Chinese Journal of Preventive Veterinary Medicine
基金 国家863项目(2011AA10A212) 黑龙江省自然科学基因项目(ZJN-0602-01)
关键词 蓝舌病病毒 VP7蛋白 NS2蛋白 单克隆抗体 bluetongue virus VP7 NS2 monoclonal antibody
分类号 S852.65 [农业科学—基础兽医学]
  • 相关文献

参考文献10

  • 1Fauquet C M, Mayo M A, Maniloff J, et al. Virus Taxonomy 8th Edition [M]. USA: Elsevier Academic Press, 2005: 466- 483.
  • 2宋红梅,杨涛,马健男,王群,张维军,徐青元,杨增岐,吴东来.蓝舌病病毒VP7基因的原核表达[J].中国预防兽医学报,2009,31(12):925-928. 被引量：10
  • 3Toussaint J F, Sailleau C, Breard E, et al. Bluetongue virus detection by two real-time RT-qPCRs targeting two different genomic segments [J]. J Virol Methods, 2007, 140(1-2): 115-123.
  • 4Mehlhom H, Walldorf V, Klimpel S, et al. First occurrence of culicoides obsoletus-transmitted bluetongue virus epidemic in central Europe [J]. Parasitol Res, 2007, 101 ( 1 ): 219- 228.
  • 5Richards R G, MacLachlan N J, Heidner H W, et al. Comparison of virologic and serologic responses of lambs and calves infected with bluetongue virus serotype 10 [J]. Vet Microbiol, 1988, 18(3-4): 233-242.
  • 6Wilson A, Mellor P. Bluetongue in Europe: vectors, epidemiology and climate change [J]. Parasitol Res, 2008, 103(I 1): 69-77.
  • 7Mann S, Mann N S, Nomikou K, et al. Novel bluetongue virus serotype from Kuwait [J]. Emerg Infect Dis, 2011, 17 (5): 886-889.
  • 8覃绍敏,白安斌,吴健敏,欧阳康,黄红梅,林俊.广西山羊蓝舌病血清学调查及流行区域分布的影响因素分析[J].中国预防兽医学报,2011,33(1):28-31. 被引量：14
  • 9Le Blois H, Fayard B, Urakawa T, et al. Synthesis and characterization of chimeric particles between epizootichemorrhagic disease virus and bluetongue vims: functional domains are conserved on the VP3 protein [J]. J Virol, 1991, 65(9): 4821.
  • 10He Cheng-qiang, Ding Nai-zheng, He Mei, et al. Intragenic recombination as a mechanism of genetic diversity in bluetongue vires [J]. J Virol, 2010, 84(21): 11487-11495.

二级参考文献18

  • 1李文超,吕茂民,杨姝,尹惠琼,史利军,马玉媛,章金刚.蓝舌病毒血清5型毒株S7基因编码区的分子克隆与序列分析[J].生物技术通讯,2007,18(3):384-387. 被引量：9
  • 2Belyaev A S, Roy P. Development of baculovirus triple and quadruple expression vectors: co-expression of three or four bluetongue virus proteins and the synthesis of bluetongue virus-like particles in insect cells [J]. Nucleic Acids Res, 1993, 21 (5): 1219-1223.
  • 3Le Blois H, Fayard B, Urakawa T, et al. Synthesis and characterization of chimeric particles between epizootic hemorrhagic disease virus and bluetongue virus: functional domains are conserved on the VP3 protein [J]. J Virol, 1991, 65(9): 4821-4831.
  • 4Loudon P T, Hirasawa T, Oldfield S, et al. Expression of the outer capsid protein VP5 of two bluetongue viruses, and sythesis of chimeric double-shelled virus-like particle using combinations of recombinant baculoviruses [J]. Virology, 1991, 182(2): 793.
  • 5Lewis S A, Grubman M J. Bluetongue virus: surface exposure of VP7 [J]. Virus Res, 1990, 16(1): 17.
  • 6Du Plessis D H, Wang L F, Jordan F A, et al. Fine mapping of a continuous epitopes on VP7 of bluetongue virus overlapping synthetic peptides and random epitopes library [J]. Virology, 1994, 198(1): 346.
  • 7Wang L F, Hyatt A D, Whiteley P L, et al. Topography and immunogenicity of bluetongue virus VP7 epitopes [J]. Arch Virol, 1996, 141(1):111.
  • 8Oldfield S, Adachi A, Urakawa T, et al. Purification and characterization of the major group-specific core antigen VP7 of bluetongue virus synthesized by a recombinant baculovirus [J]. J Gen Virol, 1990, 71: 2649-2656.
  • 9Anthony S, Jones H, Darpel K E, et al. A duplex RT-PCR assay for detection of genome segment 7 (VP7 gene) from 24 BTV serotypes [J]. J Virol Methods, 2007, 141: 188-197.
  • 10Mechamj O, Wilsonw C. Antigen capture competitive enzymelinked immunosorbent assays using baculovirus-expressed antigens for diagnosis of bluetongue virus and epizootic hemorrhagic disease virus [J]. J Clin Microbiol , 2004, 42(2): 518-523.

共引文献22

同被引文献32

  • 1林丽琴,吕敏娜,孙铭飞,廖申权,吴彩艳,戚南山,李娟,林栩慧,李国清,李华春.广东某奶牛场蓝舌病病毒分离株血清型与基因型的鉴定[J].中国兽医科学,2015,45(6):615-621. 被引量：13
  • 2Coetzee P, Stokstad M, Venter E H, et al. Bluetongue: a histori- cal and epidemiological perspective with the emphasis on South Africa [J]. J Virol, 2012, 9: 198-202.
  • 3Ratinier M, Caporale M, Golder M, et al. ldentllicatlon and characterization of a novel non-structural protein of bluetongue virus [J]. PLoS Pathogens, 2011, 7(Issue 12): e1002477.
  • 4Roy P. Bluetongue Virus Proteins [J]. J Gen Virol, 1992, 73: 3051-3064.
  • 5Roy P. Functional mapping of bluetongue virus proteins and their interactions with host proteins during virus replication [J]. Cell Biochem Biophys, 2008, 50: 143-157.
  • 6Wang Wen-shi, Sun En-cheng, Liu Ni-hong, et al. Identification of three novel linear B-cell epitopes on the VP5 protein of BTV16 [J]. Vet Microbiol, 2013, 162: 631-642.
  • 7Schwartz-Comil I,Mertens P P.Bluetongue virus:virology,pathogenesis and immunity[J].Vet Res,2008,39(5):39-46.
  • 8Vellema P.Bluetongue in sheep:Question marks on bluetongue virus serotype 8 in Europe[J].Small Ruminant Res,2008,76:141-148.
  • 9Maclachlan N J,Wilson W C,Crossley B M,et al.Novel serotype of bluetongue virus,Western North America[J].Emerg Infect Dis,2013,19(4):665-666.
  • 10Hassan S S,Roy P.Expression and functional characterization of Bluetongue virus VP2 protein:role in cell entry[J].J Virol,1999,73(12):9832-9842.

引证文献4

二级引证文献7

中国预防兽医学报
2011年 第10期

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部