Prokaryotic Expression Of SjFABP And SjGST Of Schistosoma Japonicum And Study Of Multivalent DNA Vaccine

In order to examine whether all the antigen genes inserted in multivalent DNA vaccine can be expressed in eukaryotic cells,antigen genes SjFABP and SjGST of Schistosoma japonicum were cloned and prokaryotic expressed in E.coli cells to produce recombinant antigen proteins;multivalent DNA vaccine pVIVO2-SjFABP-SjGST was constructed and expressed in mice,then using the prokaryotic expressed proteins as antigens,specific antibodies in sera were detected by western blotting.Specific primers were designed , SjFABP and SjGST genes were amplified by RT-PCR.The genes were cloned to pGEM-T Easy vectors and sequenced,then subcloned to build prokaryotic recombinants pET30a-SjFABP,pET30a-SjGST and eukaryotic recombinant pVIVO2-SjFABP-SjGST.The E.coli BL21(DE3) cells were transformed with the recombinant plasmids pET30a-SjFABP and pET30a-SjGST respectively,then induced by IPTG to express the recombinant proteins which were purified through HIS-Bind Ni-Agarose.The purified recombinant proteins were used to recognize sera from mice immunised by pVIVO2-SjFABP-SjGST.399bp and 657bp lenth PCR products were obtained;recombinant plasmids pET30a-SjFABP,pET30a-SjGST and pVIVO2-SjFABP-SjGST were constructed ,the prokaryotic expressed proteins were detected by SDS-PAGE.The rSjFABP and rSjGST can be recognized by the sera from mice immunised by pVIVO2-SjFABP-SjGST using Western blotting.The SjFABP and SjGST genes were effectively expressed in prokaryotic cells, and can be recognized by antibodies induced by DNA vaccine pVIVO2-SjFABP-SjGST.