| 犬瘟热病毒疫苗变异株的分离及其P1蛋白的表达、鉴定 |
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| 引用本文： | 于萍萍,贾红,侯绍华,袁维峰,郭晓宇,崔治中,朱鸿飞.犬瘟热病毒疫苗变异株的分离及其P1蛋白的表达、鉴定[J].中国畜牧兽医,2012,39(5):30-34. |
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| 作者姓名： | 于萍萍 贾红 侯绍华 袁维峰 郭晓宇 崔治中 朱鸿飞 |
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| 作者单位： | 1. 山东农业大学动物科技学院, 山东泰安 271017;2. 中国农业科学院北京畜牧兽医研究所, 北京 100193 |
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| 基金项目： | 公益性行业(农业)科研专项经费项目，中央级公益性科研院所基本科研业务费，“十一五”国家科技支撑计划重点项目，2011年及2012年农业部动物疫情监测与防治项目 |
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| 摘 要： | 从河北某宠物医院病犬的脾脏中分离到1株犬瘟热病毒,H基因测序结果表明,该分离株与疫苗株的同源性为99.5%,有3个氨基酸突变,但不影响其糖基化位点。将该株病毒P1基因克隆到pET32a(+)载体,转化大肠杆菌BL21(DE3)及Rosetta感受态细胞,在IPTG诱导下获得大小为50 ku,可溶性表达的P1重组蛋白。Western blotting、间接ELISA试验结果显示表达产物能被犬瘟热高免血清所识别,表明其具有良好的反应原性,有望用于犬瘟热的检测。
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| 关 键 词： | 犬瘟热病毒 变异株 P1重组蛋白 抗原性 |
Isolation of Canine Distemper Virus Vaccine Variant Strain and the Expression and Identification of P1 Recombinant Protein in E.coli |
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| YU Ping-ping
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JIA Hong
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HOU Shao-hua
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YUAN Wei-feng
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GUO Xiao-yu
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CUI Zhi-zhong
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ZHU Hong-fei.Isolation of Canine Distemper Virus Vaccine Variant Strain and the Expression and Identification of P1 Recombinant Protein in E.coli[J].China Animal Husbandry & Veterinary Medicine,2012,39(5):30-34. |
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| Authors: | YU Ping-ping JIA Hong HOU Shao-hua YUAN Wei-feng GUO Xiao-yu CUI Zhi-zhong ZHU Hong-fei |
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| Institution: | 1. Animal Science and Technology Institute, Shandong Agricultural University, Taian 271017, China;2. Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China |
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| Abstract: | A canine distemper virus was isolated from an infected dog in a pet hospital in Hebei.It had 99.5% homology with vaccine strain by sequencing of H gene,and three amino acids changed that had no effect on glycosylation sites.The P1 gene of this strain was cloned into pET32a(+) vector.The recombinant plasmid was transformed into BL21(DE3),Rosetta and protein expression was induced by IPTG.The expression protein had a molecular weight of 50 ku.And expressed mainly in form of dissolubility body.The expressed protein reacted specially with CDV serum in Western blotting and ELISA.IFA result showed that the P1 protein fixed in the cell nucleus. |
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| Keywords: | canine distemper virus variant strain P1 recombinant protein antigencity |
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