| PRRSV非结构蛋白2-半胱氨酸结构域的原核表达及抗原表位预测 |
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| 引用本文： | 周胜,高歌,孙荡,鲍梦雅,茅翔.PRRSV非结构蛋白2-半胱氨酸结构域的原核表达及抗原表位预测[J].黑龙江农业科学,2012(3):25-29. |
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| 作者姓名： | 周胜 高歌 孙荡 鲍梦雅 茅翔 |
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| 作者单位： | 南京农业大学动物医学院 |
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| 基金项目： | 南京农业大学大学生创新资助项目(1004A04) |
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| 摘 要： | 为了获得PRRSV非结构蛋白2-半胱氨酸结构域的高纯度原核表达蛋白以及了解该蛋白的抗原表位,试验以从PRRSV-VR2332毒株上提取的病毒全基因组RNA为模板,通过RT-PCR扩增非结构蛋白2-半胱氨酸结构域(Nsp2pro)基因,连接构建原核表达载体SUMO-Nsp2pro,转入宿主菌Rosetta2,经IPTG诱导,获得高浓度的可溶蛋白Nsp2pro,经亲和层析His-Bind后得到高纯度的目的蛋白。同时,运用生物信息学方法对Nsp2pro二级结构及抗原表位进行预测。结果显示,Nsp2pro的α螺旋及β折叠区域较多,转角区域较少,结构较为复杂,位于蛋白分子表面且亲水的区段很可能是B细胞表位的优势区段。获得较高纯度的蛋白,将为后续进一步研究Nsp2pro基因编码的蛋白在病毒复制过程中的作用奠定基础。
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| 关 键 词： | PRRSV Nsp2pro 基因克隆 原核表达 抗原表位预测 |
The Gene Clone,Prokaryotic Expression and Antigen Epitopes Prediction of the Cysteine Domain of Nonstructural Protein 2 of PRRSV |
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| ZHOU Sheng,GAO Ge,SUN Dang,BAO Meng-ya,MAO Xiang.The Gene Clone,Prokaryotic Expression and Antigen Epitopes Prediction of the Cysteine Domain of Nonstructural Protein 2 of PRRSV[J].Heilongjiang Agricultural Science,2012(3):25-29. |
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| Authors: | ZHOU Sheng GAO Ge SUN Dang BAO Meng-ya MAO Xiang |
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| Institution: | (Veterinary Medicine College of Nanjing Agricultural University,Nanjing,Jiangsu 210095) |
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| Abstract: | In order to acquire highly purified protein of cysteine domain of nonstructural protein 2 of PRRSV and acquaintance its antigen epiopes,the experiment were conducted with the genome extracted from PRRSV-VR2332 and got the gene of the cysteine domain of nonstructural protein 2 of PRRSV(Nsp2pro)by RT-PCR.Then,a recombinant plasmid named SUMO-Nsp2pro had been constructed,and then transformed SUMO-Nsp2pro into Rosetta2.Therefore,a large amount of Nsp2pro was obtained by IPTG.In the end,Nsp2pro was purified by His-Bind affinity chromatography,meanwhile predicted the second structure and antigen epiopes through means of bio-information,the results showed that Nsp2pro owned a great many Alpha regions and Beta regions while few of turn regions.The antigen epitopes were located in hydrolicity regions possibly.The obtained high purified protein would provide foundations for the further researches on Nsp2pro gene. |
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| Keywords: | PRRSV Nsp2pro gene clone prokaryotic expression antigen epitopes prediction |
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