Expression and identification of immune activity of cytosolic superoxide dismutase of Schistosoma japonicum
- VernacularTitle:日本血吸虫超氧化物歧化酶的表达与免疫活性鉴定
- Author:
Chuanxin YU
;
Xuren YIN
;
Jian LI
;
Yongliang XU
;
Ming XU
;
Qi GAO
;
Yousheng LIANG
- Publication Type:Journal Article
- Keywords:
Schistosoma japonicum;
Cytosolic superoxide dismutase;
Expression;
Purification;
Immunogenicity
- From:
Chinese Journal of Schistosomiasis Control
1989;0(03):-
- CountryChina
- Language:Chinese
-
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.
