| 脑心肌炎病毒VP1结构蛋白的可溶性表达及鉴定 |
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| 引用本文： | 李向茸,冯若飞,凡静静,张海霞,韦鹏建,樊得英,马忠仁. 脑心肌炎病毒VP1结构蛋白的可溶性表达及鉴定[J]. 中国兽医科学, 2011, 0(6) |
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| 作者姓名： | 李向茸 冯若飞 凡静静 张海霞 韦鹏建 樊得英 马忠仁 |
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| 作者单位： | 西北民族大学生物工程与技术国家民委重点实验室;西北民族大学生命科学与工程学院;甘肃省动物细胞工程技术研究中心; |
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| 基金项目： | 国家民委科研项目(10XB03); 甘肃省教育厅科研项目(1018B-04); 人畜共患病诊断技术平台建设校企合作项目(H2009-15) |
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| 摘 要： | 为获得可溶性表达的脑心肌炎病毒(EMCV)结构蛋白VP1片段,将EMCV VP1基因克隆到原核表达载体pGEX-4T-1中,并转化大肠杆菌BL21(DE3)表达菌中。采用不同温度、不同IPTG浓度及不同诱导时间诱导目的蛋白表达,以筛选VP1融合蛋白可溶性表达的最佳条件,并通过Western-blotting检测最佳条件下表达蛋白的免疫活性。PCR、酶切鉴定及测序结果表明,VP1基因已克隆入pGEX-4T-1质粒。经优化,确定VP1融合蛋白可溶性表达的最佳条件为:当D600nm值为0.4～0.6时,以0.25mmol/L IPTG于25℃条件下诱导表达12h。Western-blotting结果显示,诱导表达的蛋白具有免疫活性。
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| 关 键 词： | 脑心肌炎病毒 VP1基因 可溶性表达 原核表达 |
Soluble expression and identification for VP1 protein of encephalomyocarditis virus |
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| LI Xiang-rong,,FENG Ruo-fei,,FAN Jing-jing,ZHANG Hai-xia,WEI Peng-jian,FAN De-ying,MA Zhong-ren. Soluble expression and identification for VP1 protein of encephalomyocarditis virus[J]. Chinese Veterinary Science, 2011, 0(6) |
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| Authors: | LI Xiang-rong FENG Ruo-fei FAN Jing-jing ZHANG Hai-xia WEI Peng-jian FAN De-ying MA Zhong-ren |
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| Affiliation: | LI Xiang-rong1,3,FENG Ruo-fei1,2,FAN Jing-jing3,ZHANG Hai-xia3,WEI Peng-jian3,FAN De-ying1,MA Zhong-ren2(1.Key Bio-engineering and Technology Laboratory of the Nationality Commission/Northwest University for Nationalities,Lanzhou 730030,China,2.Gansu Engineering Research Center for Animal Cell,3.College of Life Science and Engineering,Northwest University for Nationalities,China) |
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| Abstract: | To express soluble VP1 in prokaryotic expression system,the gene encoding VP1 of encephalomyocarditis virus(EMCV) VP1 was cloned into the expression vector pGEX-4T-1,and the recombinant plasmid was transformed into Escherichia coli BL21(DE3).The expression conditions,including additives,IPTG concentration,temperature and induction duration were optimized.Then,Western-blotting was used to identify the immunoreactivity of the expressed protein.In result,recombinant prokaryotic expression vector pGEX-4T-1-VP1 ... |
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| Keywords: | encephalomyocarditis virus(EMCV) VP1 gene soluble expression prokaryotic expression |
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