| C型口蹄疫病毒VP1基因的原核表达及重组蛋白的纯化 |
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| 引用本文： | 常惠芸,独军政,丛国正,邵军军,林彤,冯金瑞,刘萍.C型口蹄疫病毒VP1基因的原核表达及重组蛋白的纯化[J].畜牧与兽医,2009,41(7). |
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| 作者姓名： | 常惠芸 独军政 丛国正 邵军军 林彤 冯金瑞 刘萍 |
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| 作者单位： | 1. 中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室农业部畜禽病毒学重点开放实验室,国家口蹄疫参考实验室,甘肃,兰州,730046
2. 中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室农业部畜禽病毒学重点开放实验室,国家口蹄疫参考实验室,甘肃,兰州,730046;中农威特生物科技股份有限公司,甘肃,兰州,730046 |
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| 摘 要： | 利用PCR技术扩增获得C型口蹄疫病毒VP1基因,将其克隆到原核表达载体pET-30α(+)上,构建重组表达质粒pET-VP1。质粒pET-VP1转化大肠杆菌BL21(DE3)感受态细胞,用IPTG诱导VP1基因的表达,收集不同时间的菌液进行SDS-PAGE、Western blot分析,结果得到分子量约为33 ku的目的条带,表达产物占菌体总蛋白的35%,且该目的条带能被C型FMDV阳性血清识别,说明VP1基因在大肠杆菌中得到高效表达。融合表达蛋白经镍柱纯化,重组蛋白纯度达90%以上。
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| 关 键 词： | C型口蹄疫病毒 VP1基因 表达 蛋白纯化 |
Prokaryotic expression of VP1 gene of foot-and-mouth disease virus serotype C and purification of recombinant protein |
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| CHANG Hui-yun,DU Jun-zheng,CONG Guo-zheng,SHAO Jun-jun,LIN Tong,FENG Jin-rui,LIU Ping.Prokaryotic expression of VP1 gene of foot-and-mouth disease virus serotype C and purification of recombinant protein[J].Animal Husbandry & Veterinary Medicine,2009,41(7). |
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| Authors: | CHANG Hui-yun DU Jun-zheng CONG Guo-zheng SHAO Jun-jun LIN Tong FENG Jin-rui LIU Ping |
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| Abstract: | Foot-and-mouth disease virus(FMDV) VP1 gene was amplified by RT-PCR.The VP1 fragment was inserted into plasmid pET30α(+) to obtain the recombinant plasmid pET-VP1.The pET-VP1 was transformed into Escherichia coli BL-21(DE3) and induced to express VP1 protein by IPTG.The fusion protein band with a molecular weight of 33 ku was visible on the SDS-PAGE gel.The density scanning showed that the largest amount of the fusion protein was 35% of total bacterial protein.Western blot result showed that the expression product could specifically react with the antiserum to FMDV serotype C.The fusion protein was further purified by Ni-NTA and the purified protein accounted for 90% of total protein. |
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| Keywords: | foot-and-mouth disease virus(FMDV) VP1 gene expression protein purification |
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