| O型口蹄疫病毒P1-2A基因重组杆状病毒的构建及其表达 |
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| 引用本文： | 廖德芳,信爱国,高华峰,苗海生,高林,朱明旺,李华春.O型口蹄疫病毒P1-2A基因重组杆状病毒的构建及其表达[J].中国畜牧兽医,2011,38(10):81-85. |
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| 作者姓名： | 廖德芳 信爱国 高华峰 苗海生 高林 朱明旺 李华春 |
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| 作者单位： | 1. 云南省畜牧兽医科学院,云南昆明,650224
2. 云南省保山疫苗厂,云南保山,678000 |
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| 基金项目： | 云南省应用基础研究面上项目(2009CD138); 国家公益性科研(农业)专项(201103008); 云南省农业科技创新工程项目(2008LA019) |
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| 摘 要： | 设计合成特异引物,扩增O型口蹄疫病毒(O/FMDV)P1-2A基因,将其克隆至T载体上,通过Hind Ⅲ和Not Ⅰ双酶切P1-2A基因和真核转座载体pFastBacTM Dual,构建重组转座质粒pFastBac-P12A,再将pFastBac-P12A转化入含穿梭载体Bacmid的受体菌DH10Bac,经重组筛选获得杆状病毒重组质粒Bacmid-P12A。将Bacmid-P12A质粒转染Sf9昆虫细胞,出现典型CPE。病变细胞经Dot blotting和SDS-PAGE检测和分析,结果表明,O/FMDV P1-2A蛋白在Sf9细胞中获得表达,为O型FMDV特异性蛋白。
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| 关 键 词： | 口蹄疫病毒 P1基因 重组杆状病毒 表达 |
| 收稿时间： | 2011-03-11 |
Construction and Expression of Recombinant Baculovirus with P1-2A Gene of Serotype O Foot-and-mouth Disease Virus |
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| LIAO De-fang,XIN Ai-guo,GAO Hua-feng,MIAO Hai-sheng,GAO Lin,ZHU Ming-wang,LI Hua-chun.Construction and Expression of Recombinant Baculovirus with P1-2A Gene of Serotype O Foot-and-mouth Disease Virus[J].China Animal Husbandry & Veterinary Medicine,2011,38(10):81-85. |
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| Authors: | LIAO De-fang XIN Ai-guo GAO Hua-feng MIAO Hai-sheng GAO Lin ZHU Ming-wang LI Hua-chun |
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| Institution: | 1. Yunnan Animal Science and Veterinary Institute,Kunming 650224, China;2. Yunnan Baoshan Vaccine Plant,Baoshan 678000, China |
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| Abstract: | Based on the known nucleotide sequence of FMDV gene,P1-2A gene primer was designed and synthesized.P1-2A gene was amplified by RT-PCR.The gene was then cloned into pMD18-T plasmid.The recombinant plasmids were sequenced and cloned into transfer vector pFastBacTMDual.The transfer plasmid pFastBac-P12A was constructed.pFastBac-P12A was further transferred into receptor DH10Bac bacteria containing a shuttle vector Bacmid.The recombinant plasmid Bacmid-P12A was obtained by selection,then was trans-infected into... |
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| Keywords: | FMDV P1-2A gene recombinant baculovirus expression |
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