白介素2基因与猪圆环病毒2型ORF2基因真核双表达质粒的构建

Construction of eukaryotic co-expression plasmid harboring ORF_2 gene of porcine circovirus type 2 and IL-2 gene

应用基因重组技术,构建猪圆环病毒2型ORF2基因与白介素2基因(IL-2)的真核双表达载体并将其转染CHO细胞,通过RT-PCR、间接ELISA、Western-blot、间接免疫荧光检测(IFA)等方法验证基因的表达情况。结果表明:成功构建了猪圆环病毒2型ORF2基因和IL-2基因的真核双表达质粒pIRES-ORF2-IL-2,该质粒可在体外同时表达ORF2和IL-2。

The eukaryotic co-expression plasmid pIRES-ORF2-IL-2 was constructed by inserting porcine circovirus type 2 （PCV2） ORF2 gene and IL-2 into pIRES vector using recombinant DNA technology and transfected into CHO cells. Expression of ORF2 gene of PCV2 and IL-2 in vitro was detected by RT-PCR, iELISA, Western-blot and IFA . The results indicated that co-expression plasmid harboring genes of ORF2 of PCV2 and IL-2 was successfully constructed and the plasmid could simultaneously express ORF2 gene of PCV2 and IL-2 in CHO ceils.