| 牛传染性鼻气管炎病毒gD蛋白的截短表达与活性检测 |
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| 引用本文： | 王延辉,薛飞,朱远茂,彭伍平.牛传染性鼻气管炎病毒gD蛋白的截短表达与活性检测[J].中国预防兽医学报,2007,29(11):865-869. |
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| 作者姓名： | 王延辉 薛飞 朱远茂 彭伍平 |
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| 作者单位： | 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,黑龙江,哈尔滨,150001 |
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| 基金项目： | 国家高技术研究发展计划(863计划);黑龙江省科技攻关项目 |
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| 摘 要： | 经生物学软件DNAStar分析,以牛传染性鼻气管炎病毒(Banha Nu/67)基因组DNA为模板,PCR扩增gD基因943bp的片段,将目的片段定向克隆到pET30a表达载体中,酶切及测序鉴定均正确后,转化BL21表达菌,经IPTG诱导得到部分可溶表达的重组蛋白。用Ni柱亲和层析法在非变性的条件下纯化重组蛋白,纯化的重组蛋白浓度为0.852mg/mL,纯度为85.2%。Westem blot、间接ELISA检测证明纯化的重组蛋白具有良好的抗原性和特异性。
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| 关 键 词： | 牛传染性鼻气管炎病毒 gD糖蛋白 原核表达 活性检测 |
| 文章编号： | 1008-0589（2007）11-0865-05 |
| 修稿时间： | 2007-03-08 |
Characterization of truncated glycoprotein D of infectious bovine rhinotracheitis virus expressed in E.coli |
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| WANG Yan-hui,XUE Fei,ZHU Yuan-mao,PENG Wu-ping.Characterization of truncated glycoprotein D of infectious bovine rhinotracheitis virus expressed in E.coli[J].Chinese Journal of Preventive Veterinary Medicine,2007,29(11):865-869. |
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| Authors: | WANG Yan-hui XUE Fei ZHU Yuan-mao PENG Wu-ping |
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| Institution: | National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China |
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| Abstract: | Using IBRV(Bartha Nu/67)genome DNA as the template,a 943 bp fragment of glycoprotein D gene was amplified by PCR with specific primers.The PCR product was cloned into pET30a vector and expressed in partially soluble form in E.coli aider induction of cultured E.coli with IPTG.The recombinant protein was purified by immobilized Ni ion affinity chromatography under native conditions.The purified protein was analysed by western blot and indirect enzyme-linked immunosorbent assay.The results showed that the purified recombinant protein retained good antigenicity and specificity. |
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| Keywords: | infectious bovine rhinotracheitis virus glycoprotein gD prokaryotic expression |
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