CONSTRUCTION OF DNA VACCINE OF ATP SYNTHASE LIPID-BINDING PROTEIN-LIKE PROTEIN GENE OF SCHISTOSOMA JAPONICUM AND ITS EXPRESSION IN EUKARYOTIC CELLS

Objective To clone and construct the recombinant plasmid containing ATP synthase lipid-binding protein-like protein gene of schistosoma japonicum(SjAslp)and transfer it into mammalian cells to express ATP synthase lipid-binding protein-like protein.Methods By polymerase chain reaction(PCR)technique,SjAslp was amplified from the constructed recombinant plasmid pBC SK+/SjAslp,and inserted into cloning vector pUCm-T.Then,SjAslp was subcloned into eukaryotic expression vector pcDNA3.1(+).After identifying it by PCR,restrictive enzymes digestion and DNA sequencing,the recombinant plamid was transfected into HeLa cells using electroporation,and analyzed the expression of the recombinant protein by immunocytochemistry assay.Results The specific gene fragment 558bp was successfully amplified.The DNA vaccine of SjAslp was successfully constructed.Immunocytochemistry assay showed that SjAslp was expressed in HeLa cells and located in cytoplasm.Conclusion SjAslp gene can express in eukaryotic system,which lay the foundation for development of the SjAslp DNA vaccine against schistosoma.