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兔抗猪CD163蛋白多克隆抗体制备及鉴定 被引量：1

Generation of polyclonal antibodies specifically against porcine CD163
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摘要 为制备兔抗猪CD163多克隆抗体,采用RT-PCR方法从猪PAM RNA中扩增CD163胞质外区部分基因SRCR4-5,克隆至pGEX-6p-1原核表达载体,在IPTG诱导下获得约50 000以包涵体形式表达的重组蛋白。WesternBlotting分析表明,表达产物能够被GST单抗所识别。用该重组蛋白免疫兔制备多抗血清,经间接ELISA检测,多抗血清效价可达105,流式检测发现可与Marc-145和PAM上的天然CD163分子相互作用。抗体阻断试验证明制备的多抗血清可以部分阻断PRRSV感染。本试验为进一步研究PRRSV与受体-CD163相互作用机制打下了基础。 The extracellular region of porcine CD163 gene,SRCR4-5,was amplified from total RNA of porcine alveolar macrophages and subcloned into prokaryotic expression vector pGEX-6p-1. Fol- lowed by inducing with IPTG,a 50 000 fusion protein which could be recognized by anti-GST mon- oclonal antibody was expressed. After purified,the fusion protein was employed to immunize rab- bits to generate rabbit anti-CD163 polyclonal antibodies. The polyclonal antibodies had a titer of 10s in GST-SRCR4-5 by indirect ELISA,and it also could partly block the infection of PRRSV to porcine alveolar macrophages in antibody-blocking assay. The results suggested that the polyclonal antibody was of biological function,and could be employed in the further studies on CD163 recep- tor.
作者 郭娟娟 彭金美 安同庆 冷超良 吴江 宫大庆 陈家锃 杨永倩 田志军 张德礼
机构地区 西北农林科技大学、动物医学院病毒免疫与生物信息研究室、生物信息研究中心、分子病毒免疫与肿瘤系统生物学研究组 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、猪传染病研究室
出处 《中国兽医学报》 CAS CSCD 北大核心 2012年第11期1624-1628,共5页 Chinese Journal of Veterinary Science
基金 兽医生物技术国家重点实验室开放基金重点资助项目(SKLVBF200905) 国家自然科学基金资助项目(31072115 30270342) “十二五”国家科技计划农村领域基础研究类首批预备项目(NC2010CD0178)
关键词 CD163 抗体 鉴定 CD163 antibody identification
分类号 S852 [农业科学—基础兽医学]
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参考文献15

  • 1Darvvieh L, Diaz I, Mateu E. Certainties, doubts and hypotheses in porcine reproductive and respiratory syndrome virus immunobiology[J]. Virus Res, 2010, 154(1/2) : 123-132.
  • 2Calvert J G, Slade D E, Shields S L, et al. CD163 ex- pression confers susceptibility to porcine reproductive and respiratory syndrome virus[J]. J Virol, 2007,81 (14) : 7371-7379.
  • 3Patton J B,Rowland R R,Yoo D,et al. Modulation of CD163 receptor expression and replication of porcine reproductive and respiratory syndrome virus in porcine maerophages[J]. Virus Res, 2009,140(1/2) : 161-171.
  • 4Van G H,Van B W,Delputte P L,et al. The porcine reproductive and respiratory syndrome virus requires trafficking through CD163-positive early endosomes, hut not late endosomes, for productive infection[J]. Arch Virol,2009,154(12) : 1939-1943.
  • 5Lee Y J,Park C K,Nam E,et al. Generation of a por- cine alveolar macrophage cell line for the growth of porcine reproductive and respiratory syndrome virus [J]. J Virol Methods,2010,163(2) :410-415.
  • 6Delrue I,Van G H ,Van D J. et al. Susceptible cell lines for the production of porcine reproductive and respira- tory syndrome virus by stable transfection of siaload-hesin and CD163[J]. BMC Biotechnol,2010(10):48.
  • 7Das P B,Dinh P X,Ansari I H,et al. The minor enve- lope glycoproteins GP2a and GP4 of porcine reproduc- tive and respiratory syndrome virus interact with the receptor CD163[J]. J Virol,2010,84(4) : 1731-1740.
  • 8Van G H, Van B W, Van D J, et al. Identification of the CD163 protein domains involved in infection of the porcine reproductive and respiratory syndrome virus [J]. J Virol,2010,84(6) :3101-3105.
  • 9Van B W,Delputte P L,Van G H,et al. Porcine repro- ductive and respiratory syndrome virus entry in the porcine macrophage[J ]. J Gen Viro, 2 010,91 : 16 5 9-16 6 7.
  • 10韦天超,田志军,安同庆,周艳君,肖燕,姜一峰,郝晓芳,张善瑞,彭金美,仇华吉,童光志.猪繁殖与呼吸综合征病毒Taq Man-MGB荧光定量RT-PCR方法的建立和应用[J].中国预防兽医学报,2008,30(12):944-948. 被引量：29

二级参考文献12

  • 1郭宝清,陈章水,刘文兴,崔益洙.从疑似PRRS流产胎儿分离PRRSV的研究[J].中国畜禽传染病,1996(2):1-5. 被引量：960
  • 2童光志,周艳君,郝晓芳,田志军,仇华吉,彭金美,安同庆.高致病性猪繁殖与呼吸综合征病毒的分离鉴定及其分子流行病学分析[J].中国预防兽医学报,2007,29(5):323-327. 被引量：395
  • 3Wensvoort G, Terpstra C, Pol JMA, et al. Mystery swine disease in the Netherlands: the isolation of Lelystad virus [J]. Vet Q, 1991, 13(3): 121-130.
  • 4Collins J, Benfield D A, Christianson W T, et al. Isolation of swine infertility and respiratory syndrome virus (isolate ATCC VR-2332) in North America and experimental reproduction of the disease in gnotobiotic pigs [J]. J Vet Diagn Invest, 1992, 4: 117-126.
  • 5Tian, K, Yu X, Zhao T, et al. Emergence of fatal PRRSV variants: unparalleled outbreaks of atypical PRRS in China and molecular dissection of the unique hallmark [J]. PLoS ONE, 2007, 2(6): 1-10.
  • 6Wasilk A, Callahan J D, Christopher-Hennings J, et al. Detection of U S, Lelystad, and European-Like porcine reproductive and respiratory syndrome viruses and relative quantitation in boar semen and serum samples by real-time PCR [J]. Journal of Clinical Microbiology, 2004, 42(10): 4453-4461
  • 7Chung W B, Chan W H, Chaung H C, et al. Real-time PCR for quantitation of porcine reproductive and respiratory syndrome virus and porcine circovirus type 2 in naturally-infected and challenged pigs [J]. Journal of Virological Methods, 2005, 124 (1-2): 11-19
  • 8Wills R W, Doster A R, Galeota J A, et al. Duration of infection and proportion of pigs persistently infected with porcine reproductive and respiratory syndrome virus [J]. Journal of Clinical Microbiology, 41(1): 58-62
  • 9Lew A E, Bock R E, Molloy J B, et al. Sensitive and specific detection of proviral bovine leukemia virus by 5' Taq nuclease PCR using a 3' minor groove binder fluorogenic probe [J]. J Virol Methods, 2004, 115(2): 167-175.
  • 10Johnson W, Roof M, Vaughn E, et al. Pathogenic and humoral immune responses to porcine reproductive and respiratory syndrome virus (PRRSV) are related to viral load in acute infection [J]. Veterinary Immunology and Immunopathology, 2004, 102(3): 233-247.

共引文献28

同被引文献13

  • 1Wensvoort G, Terpstra C, Pol J M A, et al. Mystery swine disease in the Netherlands: The isolation of Lelystad virus [J]. Vet Micro- biol, 1991, 13: 121-130.
  • 2Tian K, Yu X, Zhao T, et al. Emergence of fatal PRRSV variants: unparalleled outbreaks of atypical PRRS in China and molecular dissection of the unique hallmark [J]. Plos One, 2007, 2(6): 526.
  • 3Tong G Z, Zhou Y J, Hao X F, et al. Highly pathogenic porcine re- productive and respiratory syndrome, China [J]. Emerg Infect Dis, 2007, 13(9): 1434-1435.
  • 4Jusa E R, Inaba Y, Kouno M, et al. Effect of heparin on infection of cells by porcine reproductive and respiratory syndrome virus [J]. Am J Vet Res, 1997, 58 (5): 488-491.
  • 5Duma X, Nauwynck H J, Favoreel H, et al. Porcine reproductive and respiratory syndrome virus infection of alveolar macrophages can be blocked by monoclonal antibodies against cell surface anti- gens [J]. Adv Exp Med Biol, 1998, 440: 81-88.
  • 6Kim J K, Fahad A M, Shanmukhappa K, et al. Defining the cellu- lar target(s) of porcine reproductive and respiratory syndrome vi- rus blocking monoclonal antibody 7G10[J]. J Virol, 2006, 80(2): 689-696.
  • 7Calvert J G, Slade D E, Shields S L, et al. CD163 expression con- fers susceptibility to porcine reproductive and respiratory syn- drome viruses [J]. J Virol, 2007, 81(14): 7371-7379.
  • 8Zhou E M, Xiao Y, Shi Y, et al. Generation of internal image monoclonal anti-idiotypic antibodies against idiotypic antibodies to GP5 antigen of porcine reproductive and respiratory syndrome virus [J]. J Virol Methods, 2008, 149(2): 300-308.
  • 9Cao Z X, Zhao F R, Jia K, et al. Effect of compounds on the purifi- cation and antibody preparation of the extracellular domain frag- ment of the receptor CD163 [J]. Virol J, 2011, 8: 144.
  • 10马玉萍,王向鹏,赵钦,等.PRRSV细胞受体CD163真核表达及鉴定[C].广州:中国畜牧兽医学会兽医公共卫生学分会,2012:171-174.

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中国兽医学报
2012年 第11期

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