In this study,a 1.35 kb DNA fragment encoding the VP2 protein of infectious bursal disease CJ801 isolate was obtained by PCR and the sequence was determined in T vector.The VP2 gene was cloned into the vector of pcDNA...In this study,a 1.35 kb DNA fragment encoding the VP2 protein of infectious bursal disease CJ801 isolate was obtained by PCR and the sequence was determined in T vector.The VP2 gene was cloned into the vector of pcDNATM4/His/LacZ,then the expression cassette including the VP2 gene of IBDV and LacZ gene of E.coli controlled by CMV promoter were inserted into US2 gene of MDV CVI988/Rispens to give a transfer vector of pUS2-VP2.The complex of pUS2-VP2 and DOTAP was transfected into MDV infected CEF.The recombinant MDV expressing LacZ gene were selected and purified on 96-well plate by blue plague.The complete VP2 gene inserted into MDV was detected by PCR.Western blot and immunostaining results demonstrated that VP2 protein of IBDV was expressed by recombinant MDV.展开更多
文摘In this study,a 1.35 kb DNA fragment encoding the VP2 protein of infectious bursal disease CJ801 isolate was obtained by PCR and the sequence was determined in T vector.The VP2 gene was cloned into the vector of pcDNATM4/His/LacZ,then the expression cassette including the VP2 gene of IBDV and LacZ gene of E.coli controlled by CMV promoter were inserted into US2 gene of MDV CVI988/Rispens to give a transfer vector of pUS2-VP2.The complex of pUS2-VP2 and DOTAP was transfected into MDV infected CEF.The recombinant MDV expressing LacZ gene were selected and purified on 96-well plate by blue plague.The complete VP2 gene inserted into MDV was detected by PCR.Western blot and immunostaining results demonstrated that VP2 protein of IBDV was expressed by recombinant MDV.
