摘要
目的 构建Sj-FABPc表达克隆 ,获得大量纯化rSj-FABPc重组 (日本血吸虫脂肪酸结合蛋白 )抗原 ,了解其作为候选疫苗抗原的潜能。方法 以PCR法从Sj-cDNA库中扩增出目的基因片段后亚克隆入表达载体pGEX - 6P - 1进行融合表达 ,经GlutathioneSepharoseTM4B亲和层析柱和PreScissionTMProtease酶切后纯化出rSj-FABPc ,Westernblot鉴定其抗原性 ,以rSj-FABPc免疫小鼠 ,制备特异性抗血清 ,并用ELISA法检测重组抗原的免疫活性。结果 纯化的rSj-FABPc能被血吸虫成虫免疫兔血清和急、慢性血吸虫病人血清所识别 ,并能诱导小鼠产生特异性体液免疫应答 ,抗体滴度为 1∶12 80 0。结论 rSj-FABPc具有良好的抗原性 。
Aim\ To construct an expressive clone of Sj-FABPc,obtain a large amount of purified Sj-FABPc and evaluate its potency for vaccine. Methods\ The gene of Sj-FABPc was amplified by PCR, from an adult Schistosoma japonicum Chinese Mainland strain cDNA library, and subcloned into plasmid pGEX-6P-1 that codes for the 26kDa GST of Schistosoma japonicum (Sj26GST). The positive recombinant clone was induced by IPTG to express target protein which was easy to be purified with Glutathione Sepharose TM 4B and PreScission TM Protease. The purified product was analyzed by Western-blot and its immunocompetence was tested in mice and observed by ELISA. Result\ The purified rSj-FABPc can be recognized by the rabbit serum which was immunized with SWAP and the patients' serum by using western blot . The specific humoral responses was induced in mice and the titer of the specific antibody was 1:12800. Conclusion\ The rSj-FABPc has good antigenicity and can induce the immune response efficiently.
出处
《中国人兽共患病杂志》
CSCD
北大核心
2000年第5期43-45,共3页
Chinese Journal of Zoonoses
基金
江苏省重点实验室课题基金资助
关键词
日本血吸虫
脂肪酸结合蛋白
基因克隆
重组抗原
Schistosoma japonicum
Fatty acid binding protein (FABP)
Gene cloning
Recombinant antigen
Antigenicity
Immune response
