| 狂犬病病毒核蛋白在Bac-To-Bac/AcMNPV杆状病毒系统的表达 |
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| 引用本文： | 尹伟,徐洁萍,张金阳,颜焰,周继勇. 狂犬病病毒核蛋白在Bac-To-Bac/AcMNPV杆状病毒系统的表达[J]. 中国预防兽医学报, 2010, 32(2) |
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| 作者姓名： | 尹伟 徐洁萍 张金阳 颜焰 周继勇 |
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| 作者单位： | 浙江大学,农业部动物疫病病原学与免疫控制重点开放实验室,浙江,杭州,310029 |
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| 基金项目： | 国家农业公益性行业科研专项，浙江省重大科技攻关项目 |
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| 摘 要： | 为真核表达狂犬病病毒的核蛋白,本研究通过RT-PCR克隆狂犬病病毒ERA株核蛋白基因,将其克隆于杆状病毒转移载体pFastBacHTB中,构建重组质粒pFastBacHTB-NP并将其转化DH10Bac细胞,得到重组穿梭质粒reBacmid-NP;通过转染昆虫细胞sf9包装重组杆状病毒。SDS-PAGE、western blot和间接免疫荧光对表达的蛋白进行鉴定和反应原性分析。分别以重组杆状病毒表达的核蛋白、原核表达核蛋白为包被抗原进行ELISA检测。结果表明,在昆虫细胞中表达的狂犬病病毒重组核蛋白能与鼠抗RV核蛋白单克隆抗体和RV阳性血清特异性结合,其相对分子量约为50.5ku。以重组杆状病毒表达的核蛋白为抗原建立的rNP-ELISA的敏感性、特异性、符合率分别为86.36%,89.83%,90.00%,优于大肠杆菌表达的RV核蛋白。说明杆状病毒系统表达的核蛋白是建立RV核蛋白ELISA抗体检测方法的理想抗原。
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| 关 键 词： | 狂犬病病毒 核蛋白基因 Bac-To-Bac/AcMNPV杆状病毒表达系统 |
Expression of nucleoprotein of Rabies virus in Bac-To-Bac/AcMNPV Baculovirus expression system |
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| YIN Wei,XU Jie-ping,ZHANG Jin-yang,YAN Yan,ZHOU Ji-yong. Expression of nucleoprotein of Rabies virus in Bac-To-Bac/AcMNPV Baculovirus expression system[J]. Chinese Journal of Preventive Veterinary Medicine, 2010, 32(2) |
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| Authors: | YIN Wei XU Jie-ping ZHANG Jin-yang YAN Yan ZHOU Ji-yong |
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| Affiliation: | YIN Wei,XU Jie-ping,ZHANG Jin-yang,YAN Yan,ZHOU Ji-yong (Key Laboratory of Animal Epidemic Etiology , Immunological Prevention of Ministry of Agriculture,Zhejiang University,Hangzhou 310029,China) |
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| Abstract: | The nucleoprotein (NP) gene of rabies virus ERA strain was amplified by RT-PCR, cloned into the baculovirus vector pFastBacHTB and transformed into competent cells of E.coli DH10Bac. The recombinant shuttle plasmid was then transfected into sf9 insect cells. Expression of the recombinant NP (rNP) was detected by immunofluorescent assay, SDS-PAGE and western blot. The rNP had a molecular weight of 50.5 ku, and was successfully used as a coating antigen in ELISA and used the anti-NP monoclonal antibody and an... |
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| Keywords: | rabies virus nucleoprotein Bac-To-Bac/AcMNPV |
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