猪Mx1基因真核表达载体的构建及表达

Expression and construction of eukaryotic expression vector of porcine Mx1 gene

根据GenBank中猪Mx1基因序列设计并合成引物,对Mx1基因ORF区全长1 992个碱基进行扩增。将扩增片段与pVax1载体连接,构建pVa-Mx1-ORF真核表达载体。将构建的表达载体导入BHK-21和HEK-293细胞进行表达,用RT-PCR和间接免疫荧光对表达产物进行鉴定。结果表明,所构建的pVa-Mx1-ORF表达载体在BHK-21和HEK-293细胞中成功表达。

The aim of this study was to constructe the eukaryotic expression vector of porcine Mx1 gene and express the protein effectively.The open reading frame（ORF） fragment of Mx1 gene was cloned according to the sequences in Genbank,then the pVa-Mx1-ORF expression vector was constructed by linking the ORF fragment and pVax1 vector.The Mx1 protein was expressed by transforming the pVa-Mx1-ORF vector into HEK-293 and BHK-21 cells and the expression products were identified using RT-PCR and indirect immunofluorescence assay.The results revealed that the Mx1 protein was expressed successfully in HEK-293 and BHK-21 cells.This study established the foundation for research on the mechanism of Mx1 protein anti-viral infection in vitro.