›› 2013, Vol. 40 ›› Issue (3): 77-79.

• 生物技术 • 上一篇    下一篇

牛传染性鼻气管炎病毒gD蛋白主要抗原区域的高效表达、纯化与鉴定

乃德合   

  1. 青海省黄南州泽库县泽曲镇兽医站,青海黄南 811400
  • 收稿日期:2012-08-22 出版日期:2013-03-20 发布日期:2013-03-19
  • 作者简介:乃德合(1971-),男,青海人,兽医师,研究方向:传染病防治。

Truncated Expression, Purification and Identification of gD Protein of Infectious Bovine Rihinotracheitis Virus

NAI De-he   

  1. Veterinary Station of Zeku Town, Huangnan Zequ County, Qinghai Province, Huangnan 811400, China
  • Received:2012-08-22 Online:2013-03-20 Published:2013-03-19

摘要: 参照IBRV基因组序列,设计合成1对特异性引物,扩增了长约846 bp的gD基因片段。将目的片段定向克隆到pET-30a原核表达载体,酶切及测序鉴定均正确后,转化BL21(DE3)表达菌,经IPTG诱导得到了以可溶性形式表达的重组蛋白。重组蛋白纯化后,经免疫印迹检测证明具有良好的抗原性和特异性。本研究利用原核表达系统成功表达具有良好抗原性的gD蛋白,为IBRV的深入研究及诊断试剂的研制奠定了物质基础。

关键词: 牛传染性鼻气管炎病毒; gD蛋白; 原核表达; 鉴定

Abstract: According to the published IBRV genome sequence, we designed and synthetized one pair of specific primers, amplied about 846 bp gD gene fragment. The fragment was cloned into prokaryotic expression vector pET-30a, after enzyme digestion and DNA sequencing were correct, the recomninated vector was transformed into BL21(DE3) bacteria and induced by IPTG, recombinant protein was expressed in partially soluble form. After purification of recombinant protein, Western blotting proved that it had good antigenicity and specificity. The present study using prokaryotic expression system had good antigenicity of gD protein, which laid a material foundation for IBRV further research and development of diagnostic reagents.

Key words: infectious boviner rihinotracheitis virus; gD protein; prokaryotic expression; identification

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