Abstract:

This study was conducted to prepare for comparison of the prokaryotic and eukaryotic EG95-(C3d)3 proteins in the immune effects and for development of efficient method of detection Echinococcosis by building recombinant baculovirus containing three copies of C3d gene and EG95s gene of Echinococcus granulosus. EG95s gene of and three tandem copies of goat C3d 〖JP2〗gene were inserted to pTarget vector to obtain recombinant plasmid, named pTarget-EG95-(C3d)3, then the target gene, EG95-(C3d)3,were cloned into the pFastBac-Hta of Bac-to-Bac system by BamH I and Xba I, the recombinant plasmid,pFastBacHta-EG95-(C3d)3, were obtained and then〖JP〗 was transformed into the DH10Bac for transposon reorganization. The recombinant transposon, rBacmid-EG95-(C3d) 3, were transfected into Sf9 insect cells for obtain the EG95-(C3d)3 recombinant baculovirus and expression〖JP2〗 of EG95-(C3d)3 recombinant proteins. The proteins were identified by SDS-PAGE and Western blotting.rUsing the recombinant proteins detected sheep antibodies against Echinococcosis granulosa. The results showed that the EG95-(C3d)3 recombinant baculovirus was obtained, and the EG95-(C3d)3 recombinant protein with the size of about 132 ku were correctly expressed in insect cells in Sf9, the results of Western blotting showed that this EG95-(C3d)3 protein could response to Echinococcus granulosus-specific positive serum, indicating that EG95-(C3d)3 protein has immune activity. The recombinant proteins had well sensitivity with Echinococcosis granulosa. The EG95-(C3d)3 gene was correctly expressed in insect cells,which built the bases for comparison of the immune effects of prokaryotic and eukaryotic EG95-(C3d)3 proteins, and for development of efficient method of detection Echinococcosis.

Key words: Echinococcosis; EG95s gene; C3d gene; recombinant baculovirus