| 口蹄疫病毒3AB基因的克隆与原核表达 |
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| 引用本文： | 鲁会军,郑敏,韩松,李昌,金宁一.口蹄疫病毒3AB基因的克隆与原核表达[J].广西农业生物科学,2006,25(Z1):115-118. |
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| 作者姓名： | 鲁会军 郑敏 韩松 李昌 金宁一 |
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| 作者单位： | 军事医学科学院,军事兽医研究所解放军基因工程重点实验室,吉林,长春,130062 |
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| 基金项目： | 国家高技术研究发展计划(863计划) |
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| 摘 要： | 根据口蹄疫流行毒株设计一对包含3AB部分基因编码区的特异性引物,扩增出3AB基因550 bp的特异带,并将纯化的扩增产物克隆到pM D 18-T载体上。再用设计的酶切位点将3AB基因分别连接到两种原核表达载体上,构建了重组质粒pETCK S-3AB及pET 28a-3AB,转入BL 21菌中进行原核表达。SDS-PAGE电泳表明,3AB基因在两种表达系统中均高效表达。对表达的蛋白通过包涵体及切胶纯化的方法进行纯化,获得了高纯度的3AB蛋白。
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| 关 键 词： | 口蹄疫病毒 3AB基因 克隆 表达 |
| 文章编号： | 1008-3464(2006)增-0115-04 |
| 修稿时间： | 2006年8月29日 |
Cloning and prokaryotic expression of 3AB gene of FMDV |
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| LU Hui-jun,ZHENG Min,HAN Song,LI Chang,JIN Ning-yi.Cloning and prokaryotic expression of 3AB gene of FMDV[J].Journal of Guangxi Agricultural and Biological Science,2006,25(Z1):115-118. |
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| Authors: | LU Hui-jun ZHENG Min HAN Song LI Chang JIN Ning-yi |
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| Abstract: | According to the polymerase gene of FMDV,the primers of polymerase gene were designed,and the objective fragment was 550 bp which was cloned into pMD18-T vector.Then the polymerase gene was respectively cloned into two prokaryotic expression vectors,and two recombinant plasmids of pETCKS-3AB and pET28a-3AB were constructed.The plasmids were transformed to BL21(DE3) competent cell,and the polymerase was high-efficiently expressed.The two proteins were purified by the washing of inclusion bodies and gel extraction. |
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| Keywords: | FMDV 3AB gene cloning expression |
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