Construction and Expression of Eukaryotic Expression Plasmid of VP2-4-3 Gene of Very Virulent Infectious Bursal Disease Virus
Abstract: vp2-4-3 gene of very virulent infectious bursal disease virus strain SH95 (vvIBDV) was double enzyme digested and cloned into eukaryotic expression vector pALTER-MAX by cohesive ends. The recombinant plasmid was identified by enzyme-digestion, which showed that the gese was cloned into pALTER-MAX vector with right orientation and in the downstream of CMV promoter. To express this recombinant eukaryotic expression plasmid, Vero cells were transfected with the plasmids mediated by LipofectamineTM 2000 and specific proteins was detected in the cells by immuno- fluorescent assay of anti-IBDV antibody. 2-day-old chickens were intramuscularly injected with the plasmid pALTER-MAX-VP2-4-3, thigh muscles were collected at 1 week after injection. Protein expression was detected by immuno-fluorescent assay with antibody to IBDV. Expression of plasmid pALTER-MAX-V P2-4-3 in chorioallantoic membrane（CAM）of 11-day-old chicken embryo was detected by Dot-ELISA with antibody to IBDV.