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| 口蹄疫病毒L基因真核表达载体的构建及表达 | |
| 引用本文： | 刘萍,丛国正,杨孝朴,常惠芸. 口蹄疫病毒L基因真核表达载体的构建及表达[J]. 甘肃农业大学学报, 2007, 42(5): 1-4 |
| 作者姓名： | 刘萍 丛国正 杨孝朴 常惠芸 |
| 作者单位： | 1. 中国农业科学院兰州兽医研究所,甘肃,兰州,730046;甘肃农业大学动物医学院,甘肃,兰州,730070 2. 中国农业科学院兰州兽医研究所,甘肃,兰州,730046 3. 甘肃农业大学动物医学院,甘肃,兰州,730070 |
| 基金项目： | 国家重点基础研究发展计划(973计划) |
| 摘 要： | 采用RT-PCR方法,从口蹄疫病毒中扩增出长约600 bp的核苷酸片段.纯化后与载体pMD18-Tsimple连接,重组质粒经PCR鉴定及DNA测序,结果表明克隆的片段为口蹄疫病毒L基因.将质粒pMD18-L与表达载体pEGFP-N1分别用BamHⅠ+XhoLⅠ双酶切,将所获目的基因与带有酶切位点的载体连接,经酶切、PCR鉴定及DNA测序,结果表明重组表达质粒pEGFP-L构建成功.将转染的BHK-21细胞,经荧光鉴定和RT-PCR检测,证实L基因在BHK-21细胞中得到表达. |
| 关 键 词： | 口蹄疫病毒 L基因 真核表达 |
| 文章编号： | 1003-4315（2007）05-0001-04 |
| 修稿时间： | 2006-10-10 |
Construction of eukaryotic expressing vectors of L gene of foot-and-mouth disease virus |
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| LIU Ping,CONG Guo-zheng,YANG Xiao-pu,CHANG Hui-yun. Construction of eukaryotic expressing vectors of L gene of foot-and-mouth disease virus[J]. Journal of Gansu Agricultural University, 2007, 42(5): 1-4 | |
| Authors: | LIU Ping CONG Guo-zheng YANG Xiao-pu CHANG Hui-yun |
| Affiliation: | 1. Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China; 2. College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070 ,China |
| Abstract: | The L gene of foot-and-mouth disease virus was amplified by RT-PCR,the purified DNA fragment was inserted into pMD18-T vector,which contains complete L gene.The plasmid pMD18-L and the expression vector pEGFP-N1 was digested by BamH I XhoL I respectively,and the amplicon was cloned into pEGFP-N1 to construct recombinant expression plasmid.The recombinant plasmid was checked by restriction enzyme,PCR analysis and nucleic acid sequencing.Further BHK-21 cells were transfected with pEGFP-L.The expression proteins were analyzed by fluoroscopy and RT-PCR.The results showed that proteins of L was expressed in BHK-21 cells. |
| Keywords: | FMDV L gene eukaryotic expression |
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