Construction and expression of eukaryotic expression plasmid for F gene of canine distemper virus Isolated from mink

In order to construct eukaryotic expression plasmid for F gene of canine distemper virus,the F gene of CDV strain isolated from mink was amplified by reverse transcription polymerase chain reaction(RT-PCR).The amplified fragments were cloned into pMD-18T simple vector.Objective gene was obtained by digesting the cloning plasmid with BamHⅠand KpnⅠ,and the eukaryon recombinant expression plasmids was constructed by re-cloning the objective gene into pcDNA3.1(+) vector.Eukaryon recombinant expression plasmid pcDNA3.1-CDVF was transfected into BHK-21 cell by using lipoplast mediate method and it was proved with indirect fluorescent antibody technique and RT-PCR detection that the objective gene was successfully expressed in BHK-21 cell.The results of the study provided fundamental information for the further study on the prevention of mink CD and new generation vaccine.