| 波形蛋白介导猪繁殖与呼吸综合征病毒感染Marc-145细胞作用的初步研究 |
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| 引用本文： | 王伟伟,张璐,马晓春,高继明,肖一红,周恩民.波形蛋白介导猪繁殖与呼吸综合征病毒感染Marc-145细胞作用的初步研究[J].病毒学报,2011,27(5):456-461. |
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| 作者姓名： | 王伟伟 张璐 马晓春 高继明 肖一红 周恩民 |
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| 作者单位： | 山东农业大学动物医学院,山东泰安,271018;西北农林科技大学动物医学院,陕西杨凌,712100 |
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| 基金项目： | 国家自然科学基金(30871857,30901063,U0931003/L01); 高等学校博士学科点专项科研基金(20093702120004) |
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| 摘 要： | 为了研究波形蛋白在介导PRRSV感染过程中的作用,根据GenBank中已发表的波形蛋白序列,设计特异性引物,利用RT-PCR方法从Marc-145细胞中扩增目的基因,克隆入pET-28a,转化表达菌BL21(DE3)中进行诱导表达,用SDS-PAGE和Western blot鉴定表达产物,将表达产物纯化后免疫BALB/c小鼠制备血清,用病毒阻断实验验证PRRSV与波形蛋白及其抗体的关系。用ELISA方法确定了重组波形蛋白与PRRSV结构蛋白N蛋白和囊膜蛋白GP5蛋白之间的关系。结果表明,成功的从Marc-145细胞中扩增出全长的波形蛋白基因,将其克隆入pET-28a,诱导后得到高效表达,表达蛋白纯化后免疫小鼠抗体效价达到105,病毒阻断实验表明波形蛋白能部分阻断PRRSV感染,其抗血清能完全阻断PRRSV感染。ELISA结果表明波形蛋白能与N蛋白结合而不与GP5蛋白结合。此结果为PRRSV感染的细胞的受体机制增添了新的内容,为PRRSV感染过程中受体间的相互作用关系研究奠定基础。
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| 关 键 词： | 猪繁殖与呼吸综合征病毒 波形蛋白 表达 病毒阻断 N蛋白 |
The role of vimentin during PRRSV infection of Marc-145 cells |
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| Wang Wei-Wei,Zhang Lu,Ma Xiao-Chun,Gao Ji-Ming,Xiao Yi-Hong,Zhou En-Min.The role of vimentin during PRRSV infection of Marc-145 cells[J].Chinese Journal of Virology,2011,27(5):456-461. |
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| Authors: | Wang Wei-Wei Zhang Lu Ma Xiao-Chun Gao Ji-Ming Xiao Yi-Hong Zhou En-Min |
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| Institution: | WANG Wei-wei1,ZHANG Lu1,MA Xiao-chun1,GAO Ji-ming1,XIAO Yi-hong1,ZHOU En-min2 (1.College of Animal Science and Veterinary Medicine,Shandong Agricultural University,TaiAn,Shandong 271018,China,2.College of Veterinary Medicine,Northwest A & F University,Yangling,Shaanxi 712100,China) |
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| Abstract: | The objective of this study was to investigate the function of vimentin in PRRSV infection.Vimentin gene from Marc-145 cells was amplified by RT-PCR,cloned into pET-28a vector and expressed in Escherichia coli BL21(DE3).The expressed vimentin was confirmed by Western blot and purified which was used to immunize BALB/c mice for the production of antibodies.Vimentin and antibodies were tested for blocking PRRSV infection of Marc-145 cells.The binding of vimentin to PRRSV N and GP5 proteins were tested by the ... |
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| Keywords: | PRRSV vimentin virus blocking N protein |
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