Construction and identification of a recombinant Bifidobacterium bifidum(pGEX-Sj26GST-Sj32) vaccine against Schistosoma japonicum

Objective To construct and identify a recombinant Bifidobacterium bifidum(Bb)(pGEX-Sj26GST-Sj32) vaccine against Schistosoma japonicum(Sj).Methods The plasmid pET28α-Sj26GST-Sj32 was extracted from recombinant BL21(DE3)(pET28α-Sj26GST-Sj32) stored in our laboratory.The fusion gene Sj26GST-Sj32 was amplified with PCR and served as a template for cloning into Escherichia coli-Bifidobacteria shuttle expression vector pGEX-1λT to construct recombinant plasmid pGEX-Sj26GST-Sj32.The recombinant plasmid was transformed into E.coli BL21(DE3) and was extracted and identified.BamHⅠ and EcoRⅠ were used to identify the length of the vector fragment and target gene fragment.pGEX-Sj26GST-Sj32 was then electroporated into Bb to construct a recombinant Bb(pGEX-Sj26GST-Sj32) vaccine.The plasmid pGEX-Sj26GST-Sj32 of recombinant Bb was extracted and identified using PCR.Results According to PCR,the Sj26GST-Sj32 fusion gene was 1 991 bp in length.The electrophoretogram of the products digested by BamHⅠ and EcoRⅠ revealed that the Sj26GST-Sj32 fusion gene was cloned into the plasmid pGEX-1λT.PCR revealed that the fusion gene Sj26GST-Sj32 1 991 bp in length was amplified from the template of the recombinant Bb(pGEX-Sj26GST-Sj32) vaccine.Conclusion A recombinant Bb(pGEX-Sj26GST-Sj32) vaccine against Sj was successfully constructed.