Expression and identification of immune activity of cytosolic superoxide dismutase of Schistosoma japonicum / 中国血吸虫病防治杂志

ABSTRACT

Objective To express the cytosolic superoxide dismutase(SOD)of Schistosoma japonicum and analyze its antigenicity.MethodsThe DNA sequence of Schistosoma japonicum gene of cytosolic SOD was amplified by reverse transcription polymerase chain reaction(RT-PCR)and subcloned into the expression vector pGEX-4T-3 to construct a recombinant plasmid Sj SOD-pGEX-4T-3.This recombinant plasmid was transformed into component cell of E.coli BL21(DE3).The fusion protein of GST-SOD was expressed by inducing with IPTG and purification by affinity chromatography.The specific antiserum was prepared by immunizing the BALB/c mouse with GST-SOD fusion protein,and the immnuogenicity of GST-SOD was investigated by Western blot analysis.ResultsThe gene of cytosolic SOD was amplified successfully and subcloned into expression vector pGEX-4T-3 forming the recombinant expression plasmid Sj SOD-pGEX-4T-3.The fusion protein GST-SOD was expressed after the recombinant containing Sj SOD-pGEX-4T-3 being induced by IPTG.Immunizing the BALB/c mouse with the fusion protein GST-SOD produced high titer specific antiserum of 1∶12800 and the fusion protein GST-SOD was recognized by sera of severe infection rabbits.ConclusionsThe cytosolic SOD has been expressed successfully and has a preferable immunogenicity.