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当前位置: 首页 > 文章 > O型口蹄疫病毒P1-2A基因重组杆状病毒的构建及其表达 中国畜牧兽医 2011,38 (10) 83-87
Position: Home > Articles > Construction and Expression of Recombinant Baculovirus with P1-2A Gene of Serotype O Foot-and-mouth Disease Virus China Animal Husbandry & Veterinary Medicine 2011,38 (10) 83-87

O型口蹄疫病毒P1-2A基因重组杆状病毒的构建及其表达

作  者：
廖德芳;信爱国;高华峰;苗海生;高林;朱明旺;李华春
单  位：
云南省保山疫苗厂;云南省畜牧兽医科学院
关键词：
口蹄疫病毒;P1基因;重组杆状病毒;表达
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HindⅢ和NotⅠ双酶切P1-2A基因和真核转座载体pFastBacTMDual,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。
译  名：
Construction and Expression of Recombinant Baculovirus with P1-2A Gene of Serotype O Foot-and-mouth Disease Virus
作  者：
LIAO De-fang1,XIN Ai-guo1,GAO Hua-feng1,MIAO Hai-sheng1,GAO Lin1,ZHU Ming-wang2,LI Hua-chun1(1.Yunnan Animal Science and Veterinary Institute,Kunming 650224,China; 2.Yunnan Baoshan Vaccine Plant,Baoshan 678000,China)
关键词：
FMDV;P1-2A gene;recombinant baculovirus;expression
摘  要：
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 Sf9 insect cells.The recombinant baculovirus which expressing serotype O FMDV P1-2A gene was harvested.The Sf9 cells were trans-infected with recombinant baculovirus expressing serotype O FMDV P1-2A gene and showed typical CPE.The cells were harvested and tested by FMDV Dot blotting and SDS-PAGE analysis.Results indicated that the serotype O FMDV P1-2A gene expressed in Sf9 cells and the P1-2A protein antigen was specific to serotype O FMDV.

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