日本血吸虫(中国大陆株)线粒体相关的重组融合蛋白(rSj338/26GST)的免疫原性鉴定  

IMMUNOGENICITY IDENTIFICATION OF RECOMBINANT MITOCHONDRIA-RELATED PROTEIN OF SCHISTOSOMA JAPONICUM (CHINESE STRAIN)

作　　者：李春玲[1] 胡雪梅[2] 张兆松[1] 季旻珺[1] 苏川[1] 吴海玮[1] 

机构地区：[1]南京医科大学医学分子生物学研究所分子免疫寄生虫学研究室,南京210029 [2]山东滨州医学院病原生物学教研室

出　　处：《中国血吸虫病防治杂志》2002年第3期184-187,共4页Chinese Journal of Schistosomiasis Control

基　　金：国家自然科学基金 ( 3980 0 316 );总理预备金血防专项基金 ( 94 - y- 2 3) (部分资助 )

摘　　要：目的　鉴定纯化的重组日本血吸虫线粒体相关融合蛋白 (r Sj3 3 8/2 6GST)的免疫原性。方法　对已构建的阳性表达菌 p GEX-6p-1/Sj3 3 8进行大量诱导表达 ,并对纯化蛋白进行活性分析。将粗提包涵体蛋白、复性包涵体蛋白、经分子筛纯化并复性的蛋白分别免疫新西兰家兔 2只。用 West-ern blot及 Dot-EL ISA方法检测特异性抗体的免疫原性及各组抗体水平。结果　用分子筛纯化的融合蛋白 r Sj3 3 8/2 6GST经 SDS-PAGE电泳鉴定显示达电泳纯 ;Western blot显示纯化蛋白能够被日本血吸虫重感染兔血清及 r Sj3 3 8/2 6GST免疫兔血清识别。Dot-ELISA法检测结果表明 ,经分子筛纯化并复性的 r Sj3 3 8/2 6GST融合蛋白免疫兔血清中特异性抗体的滴度最高 ,为 1∶ 5 12 0 0。结论　纯化后的融合蛋白 r Sj3 3 8/2 6GST具有较高的纯度及较强的免疫活性 ,可望作为日本血吸虫病疫苗候选分子 。Objective To identify the immunogenicity of the purified S.japonicum mitochondria related protein rSj338/26GST.Methods The positive expression clone pGES 6p 1/Sj338 was induced by IPTG to produce sufficient recombinant fusion protein rSj338/26GST. The inclusion bodies, the revivified inclusion bodies and the purified protein from S 12 gel filtration were analyzed with Western blot and then injected into two rabbits, respectively. The immunogenicity of the specific anti Sj338/26GST antibody was identified by Western blot and its level was analyzed by Dot ELISA. Results The inclusion body, the revivified inclusion bodied and the purified protein from S 12 gel filtration could be recognized by the sera from the rabbits heavily infected with S.japonicum and its SWAP immunized rabbit sera. The level of the specific antibody was the highest in those rabbits which were immunized with the revivified protein and purified protein from S 12 gel filtration. Conclusion The purified fusion protein Sj338/26GST has higher immunogenicity and may be a new vaccine candidate.

关 键 词：日本血吸虫 线粒体 重组融合蛋白 rSj338/26GST 重组抗原 免疫原性 

分 类 号：R532.21[医药卫生—内科学] R383.24[医药卫生—临床医学]