O型口蹄疫病毒P1-2A基因重组杆状病毒的构建及其表达  

Construction and Expression of Recombinant Baculovirus with P1-2A Gene of Serotype O Foot-and-mouth Disease Virus

作　　者：廖德芳 信爱国 高华峰 苗海生 高林 朱明旺 李华春 

机构地区：[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

出　　处：《中国畜牧兽医》2011年第10期81-85,共5页China Animal Husbandry & Veterinary Medicine

基　　金：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)

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。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 pFastBacTM Dual. 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.

关 键 词：口蹄疫病毒 P1基因 重组杆状病毒 表达 

分 类 号：Q78[生物学—分子生物学]