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| 口蹄疫病毒非结构蛋白2C基因在昆虫细胞中的表达及应用 | |
| 引用本文： | 张小丽,卢曾军,马小军,曹轶梅,付元芳,刘在新,谢庆阁.口蹄疫病毒非结构蛋白2C基因在昆虫细胞中的表达及应用[J].畜牧兽医学报,2011,42(12). |
| 作者姓名： | 张小丽 卢曾军 马小军 曹轶梅 付元芳 刘在新 谢庆阁 |
| 作者单位： | 1. 甘肃农业大学动物医学院,兰州730070;家畜疫病病原生物学国家重点实验室,国家口蹄疫参考实验室,中国农业科学院兰州兽医研究所,兰州730046 2. 家畜疫病病原生物学国家重点实验室,国家口蹄疫参考实验室,中国农业科学院兰州兽医研究所,兰州730046 3. 甘肃农业大学动物医学院,兰州,730070 |
| 基金项目： | 国家支撑计划(2006BAD06A12); 国家“973”计划(2005CB523201) |
| 摘 要： | 研究口蹄疫病毒(FMDV)非结构蛋白(NSP) 2C在区分灭活疫苗免疫动物与自然感染动物方面的意义.本研究将FMDV NSP 2C基因,克隆到穿梭载体pFast-bac- HT-B,将其转入含骨架载体Bacmid的DH10Bac,经蓝白斑筛选得到重组骨架质粒Bacmid-2C.将Bacmid-2C转染昆虫细胞Sf9,鉴定正确后,经3次增殖获得高滴度的P-3代病毒后,在High Five细胞中进行目的蛋白的表达.SDS-PAGE结果显示在High Five细胞中得到了相对分子质量约为38.93 ku的目的蛋白2C,Western blotting及Dot-ELISA结果显示,该表达产物对FMDV感染动物阳性血清有良好的反应性.以电泳纯化的2C蛋白为抗原建立间接ELISA,检测健康非免疫动物、免疫动物及FM-DV试验感染动物血清,结果表明2C-ELISA不但能区分免疫动物和感染动物的血清,而且还能检测FMDV感染早期动物血清中的2C抗体.说明昆虫细胞表达的2C蛋白可作为FMDV疫苗免疫动物与自然感染动物鉴别诊断的良好抗原.2C基因在Bac-to-Bac系统中的成功表达为建立通过检测几种NSP抗体,筛查感染及隐性带毒动物,净化畜群的方法奠定了基础. |
| 关 键 词： | 口蹄疫病毒 2C基因 昆虫杆状病毒 表达 应用 |
Expression of Non-Structural Protein 2C of Foot-and-mouth Disease Virus in Insect Cells and Its Application |
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| ZHANG Xiao-li , LU Zeng-jun , MA Xiao-jun , CAO Yi-mei , FU Yuan-fan , LIU Zai-xin , XIE Qing-ge.Expression of Non-Structural Protein 2C of Foot-and-mouth Disease Virus in Insect Cells and Its Application[J].Acta Veterinaria et Zootechnica Sinica,2011,42(12). | |
| Authors: | ZHANG Xiao-li LU Zeng-jun MA Xiao-jun CAO Yi-mei FU Yuan-fan LIU Zai-xin XIE Qing-ge |
| Institution: | ZHANG Xiao-li1,2,LU Zeng-jun2,MA Xiao-jun1,CAO Yi-mei2,FU Yuan-fang2,LIU Zai-xin2,XIE Qing-ge2 (1.College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China,2.State Key Laboratory of Veterinary Etiological Biology,National Foot-and-mouth Disease Reference Laboratory,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China) |
| Abstract: | In recent years,the potential value of nonstructural protein(NSP) 2C was well documented for distinguishing foot-and-mouth disease virus(FMDV) infected animals from vaccinated animals.In this study,2C gene of FMDV was cloned into baculovirus transfer vector pFast-bac-HT-B,and then the recombinant vectors were transformed into DH10Bac which contain backbone Bacmid.The white colonies were selected and Bacmid-2C recombinant vectors were used for transfection of Sf9 cells.After three times amplification of the ... |
| Keywords: | foot-and-mouth disease virus NSP 2C insect cell expression application |
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