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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (11): 86-88.doi: 10.11924/j.issn.1000-6850.2011-3441

所属专题： 生物技术； 畜牧兽医

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

兔出血症病毒VP60基因的表达及其免疫原性研究

隋慧   

  • 收稿日期:2011-11-18 修回日期:2011-12-06 出版日期:2012-04-15 发布日期:2012-04-15

Expression and Immunogenicity of VP60 Gene of Rabbit Haemorrhagic Disease Virus

  • Received:2011-11-18 Revised:2011-12-06 Online:2012-04-15 Published:2012-04-15

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摘要：

为研究兔出血症病毒VP60基因原核表达蛋白的免疫原性。采用RT-PCR方法扩增了RHDV衣壳蛋白VP60基因。PCR产物纯化后连接T载体，提取质粒后进行序列测定，将测序正确的纯化产物经双酶切，克隆入原核表达载体PET28b中，构建原核表达载体。将鉴定正确的重组质粒转化表达宿主菌E. coli RosettaTM中，用IPTG诱导培养重组表达菌。结果表明：经SDS-PAGE分析，在相对分子量62.0 KD处可见明显的表达带，经Western blot鉴定，在62.0 KD处出现明显的反应带。以纯化的重组蛋白制备的抗血清可以与纯化的RHDV发生特异性的ELISA反应。说明RHDVVP60基因原核表达蛋白具有较好的免疫原性。

关键词: 绥化市, 绥化市

Abstract:

The aim of this article was to investigate the immunogenicity of rabbit haemorrhagic disease virus (RHDV) VP60 gene which was expressed in a prokaryotic system. The capsid protein VP60 gene of RHDV was cloned by RT-PCR and the product after purification was linked into PMD-18T vector. The plasmid was sequenced after extraction. Then the amply product which was digested by dual-enzyme was linked into prokaryotic expression vector PET28b and constructed prokaryotic expression vector. The recombinant plasmid was transformed into host strain E. coli RosettaTM induced by IPTG after the identification was right. The results showed that the expressed band of 62.0 KD was identified by SDS-PAGE and western blot. The purified recombinant protein reacted with the purified RHDV in ELISA. The prokaryotically expressed protein of RHDV VP60 showed well immunogenicity.