PRRSV GP5和PCV2 ORF2双基因共表达重组腺病毒的构建及鉴定  

Construction of recombinant adenovirus co-expressing PRRSV GP5 gene and PCV2 ORF2 gene

作　　者：于新友 沈志强[1,2] 管宇[2] 南松剑[2] 王金良[2] 肖跃强[2] 唐娜[2] 

机构地区：[1]山东省畜禽用蜂胶疫苗工程技术研究中心,山东滨州256600 [2]山东省滨州畜牧兽医研究院,山东滨州256600

出　　处：《中国预防兽医学报》2009年第9期667-670,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：山东省自然科学基金项目(Y2006D26)

摘　　要：为构建猪繁殖与呼吸综合征病毒(PRRSV)GP5基因和猪圆环病毒2(PCV2)ORF2双基因共表达重组腺病毒。将扩增的PRRSV GP5基因和PCV2 ORF2基因通过IRES元件串联,构建重组腺病毒穿梭质粒。PmeⅠ酶切线性化后,转化含有腺病毒骨架载体pAdeasy-1的大肠杆菌BJ5183感受态细胞,同源重组后筛选重组腺病毒质粒,经PacⅠ酶切后转染293A细胞进行包装,获得重组腺病毒。PCR鉴定表明重组腺病毒含有GP5和ORF2基因,western blot检测结果表明GP5基因和ORF2基因在293A细胞内获得表达。To construct recombinant adnovirus that expresses the PRRSV GP5 and PCV20RF2 proteins, recombinant shuttle plasmid was constructed by linking the PRRSV GP5 and PCV20RF2 genes via IRES gene. The plasmid was linerized with Pme I and transformed into Escherichia coli B J5183 competent cells to allow homologous recombination with the supercoiled adenoviral vector pAdEasy-1. The recombinant adenovirus plasmid was digested with Pac I and transfected into 293 A cells to pack into viral particles, The recombinant adenovirus containing GP5 and PCV2 gene was identified by PCR. The western blot results showed that the GP5 gene of PRRSV and ORF2 gene of PCV2 was successfully expressed in 293 A cells.

关 键 词：猪繁殖与呼吸综合征病毒(PRRSV) GP5基因 猪圆环病毒2型(PCV2) ORF2基因 重组腺病毒 

分 类 号：S852.65[农业科学—基础兽医学]