摘要
选取O型口蹄疫病毒(FMDV)OMⅢ株的P1-2A和3C作为目的基因,分别扩增出目的片断后,定向亚克隆至腺病毒穿梭质粒pAdTrack-CMV中,得到重组载体pAdTrack-P12A3C,重组穿梭质粒和腺病毒骨架载体pAdeasy-1共同电转化入大肠杆菌BJ5183感受态细胞,同源重组后得到重组腺病毒质粒(pAd-P12A3C)。将重组腺病毒质粒线性化后转染至HEK293细胞中可观察到典型的绿色荧光,从重组腺病毒的基因组中可扩增到P12A3C基因,证实获得了含有P12A3C的重组腺病毒。
To construct recombinant adenovirus containing the P1-2A-3C genes of FMDV serotype OMⅢ,the genes of P1-2A-3C were obtained by RT-PCR.The amplification products were digested with double restriction endonuclease respectively,and orientationally subcloned into shuttle vector pAdTrack-CMV.The recombinant shuttle plasmid was named pAdTrack-P12A3C,and then the recombinant adenoviral plasmid was obtained by homologous recombination of the linearized pAdTrack-P12A3C and pAdEasy-1 vector into E.coli strain BJ5183,finally the recombinant adenoviral plasmid was cleaved with PacI and transfected into HEK293 packaging cell line.The green fluorescence can be seen in infected HEK293 cells and P1-2A-3C genes can be amplified in recombinant adenoviral genome,those results demonstrate the strain of reco mbinant adenovirus containing P1-2A-3C genes from the FMDV was successfully obtained.
出处
《湖南农业科学》
2010年第6期125-127,130,共4页
Hunan Agricultural Sciences
关键词
口蹄疫病毒
P12A3C基因
重组腺病毒
foot-and-mouth disease virus（FMDV）
P1-2A-3C genes
recombinant adenovirus
