番鸭呼肠孤病毒σC基因重组腺病毒载体的构建和鉴定  

Construction and Certification of Recombinant Adenovirus Vector for σC Gene in Muscovy Duck Reovirus

作　　者：林锋强[1] 程晓霞[1] 王劭[1] 朱小丽[1] 王锦祥[1] 陈仕龙[1] 陈少莺[1] 

机构地区：[1]福建省农业科学院畜牧兽医研究所/福建省畜禽疫病防治工程技术研究中心,福建福州350013

出　　处：《福建农业学报》2016年第10期1015-1019,共5页Fujian Journal of Agricultural Sciences

基　　金：国家自然科学基金项目(31172334);福建省自然科学基金项目(2016J01137)

摘　　要：应用RT-PCR方法扩增出番鸭呼肠孤病毒(Muscovy duck reovirus,MDRV)的σC基因,通过XhoⅠ+HindⅢ双酶切后把该基因插入到经过同样双酶切的穿梭质粒pShuttle-CMV载体中。重组穿梭载体经过双酶切和PCR鉴定后进行测序,序列测定正确,同源性为99.8%。将获得的重组穿梭质粒与腺病毒骨架质粒共转染293细胞后获得重组腺病毒。PCR和RT-PCR鉴定的结果证明σC蛋白基因已成功插入到重组腺病毒载体中。The aC gene of muscovy duck reovirus （MDRV） was amplified by RT-PCR and cloned into pShuttle-CMV plasmid after the amplicon was digested by XhoI＋ Hind III enzyme. Sequencing analysis confirmed the gene to be aC. Subsequently, the recombinant plasmid pShuttle-CMV-σC and the adenovirus backbone vector, pAdEasy-1, were co-transfected into 293 cells to construct a recombinant adenovirus. The obtained recombinant adenovirus pAd- σC was positively identified by PCR and RT-PCR. The information obtained could be used for genetic engineering a vaccine against MDRV.

关 键 词：番鸭呼肠孤病毒 σC基因 重组腺病毒载体 

分 类 号：S852[农业科学—基础兽医学]