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日本血吸虫还原性辅酶I(SjNADH)基因的克隆表达及其免疫保护效果分析
引用本文:赵彬,洪炀,曹燕,韩艳辉,吴秀娟,石耀军,林矫矫,陈铁桥,傅志强.日本血吸虫还原性辅酶I(SjNADH)基因的克隆表达及其免疫保护效果分析[J].中国动物传染病学报,2012(2):59-64.
作者姓名:赵彬  洪炀  曹燕  韩艳辉  吴秀娟  石耀军  林矫矫  陈铁桥  傅志强
作者单位:湖南农业大学动物医学院;中国农业科学院上海兽医研究所 农业部动物寄生虫学重点开放实验室
基金项目:公益性行业(农业)科研专项(200903036);基本科研业务费(2010JB10)
摘    要:为了评估日本血吸虫还原性辅酶I(SjNADH)在小鼠体内诱导的免疫保护效果,应用PCR获得SjNADH基因片段,其开放阅读框为474 bp,编码157个氨基酸,并在大肠杆菌中成功表达,纯化得到该重组蛋白。Western blot结果显示,SjNADH在日本血吸虫童虫和成虫中的表达量稳定,SjNADH融合蛋白也能被免疫血清识别,具有较好的免疫原性。应用重组蛋白免疫小鼠能诱导产生较高的特异性抗体水平,并诱导了20.13%的减虫率和23.06%的肝脏减卵率。结果表明,获得的SjNADH重组蛋白在小鼠体内诱导产生了部分免疫保护,有作为血吸虫疫苗候选抗原分子的潜力。

关 键 词:日本血吸虫  还原性辅酶I  克隆  表达  免疫保护

CLONING,EXPRESSION AND IMMUNOGENICITY OF SCHISTOSOMA JAPONICUM NADH I
ZHAO Bin,HONG Yang,CAO Yan,HAN Yan-hui,WU Xiu-jun,SHI Yao-juan,LIN Jiao-jiao,CHEN Tie-qiao,FU Zhi-qiang.CLONING,EXPRESSION AND IMMUNOGENICITY OF SCHISTOSOMA JAPONICUM NADH I[J].Chinese Journal of Animal Infectious Diseases,2012(2):59-64.
Authors:ZHAO Bin  HONG Yang  CAO Yan  HAN Yan-hui  WU Xiu-jun  SHI Yao-juan  LIN Jiao-jiao  CHEN Tie-qiao  FU Zhi-qiang
Institution:1.Hunan Agricultural University Veterinary Faculty,Changsha 410128,China;2.Key Laboratory of Animal Parasitology,Ministry of Agriculture,Shanghai Veterinary Reserch Institute,CAAS,Shanghai 200241,China)
Abstract:The objectives of the present study were to clone and express the gene encoding NADH I of Schistosoma japonicum and evaluate its immunoprotection activity in mice.Sequence analysis showed that the ORF of SjNADH was 474 bp,encoding 157 amino acids.The recombinant protein was purified from E.coil BL21(DE3) with His·Bind resin,and showed good immunogenicity in mice.The contents of SjNADH in schistosomula and adult worm were similar as determined in Western blot.Mice vaccinated with recombinant protein showed a high level of specific antibody response,worm burden reduction rate of 20.13% and liver egg burden reduction of 23.06%.Results suggested that the recombinant pET28a(+)-SjNADH protein had the potential as a vaccine candidate against schistosomiasis.
Keywords:Schistosoma japonicum  NADH  clone  expression  immunoprotection
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