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日本血吸虫22.6kDa抗原T细胞表位的重组、表达及初步鉴定 被引量:7

Preliminary Identification of T Cell Epitopes on 22.6kDa Antigen of Schistosoma japonicum
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摘要 目的 鉴定日本血吸虫中国大陆株 2 2 .6kDa抗原 (Sj2 2 .6 )的T细胞表位。 方法 用计算机软件预测Sj2 2 .6分子的T细胞表位 ,设计并合成其编码核苷酸 ,定向克隆入融合表达载体pET 32c( + ) ,转化大肠杆菌BL2 1感受态细胞 ,经酶切及测序鉴定出重组克隆。阳性克隆经IPTG诱导表达 ,表达产物用NTA柱纯化。用纯化后的表位肽融合蛋白体外刺激C3H小鼠脾单个核细胞 ,3 H TdR掺入法检测其增殖。 结果 用计算机软件预测获得Sj2 2 .6抗原的 4个候选表位肽 ,并成功克隆入pET 32c( + )。其重组体均可表达分子量约 2 0kDa的融合蛋白 ,用NTA柱纯化后经 1 2 %聚丙烯酰胺凝胶电泳 (SDS PAGE)显示单一条带。其中P4、P5、P6融合蛋白能有效刺激小鼠脾单个核细胞增殖。 结论 从Sj2 2 .6抗原中初步鉴定出P4、P5、P6 Objective To identify the T cell epitopes on 22.6kDa antigen of Schistosoma japonicum (Sj22.6). Methods The primary structure of Sj22.6 molecule was analysed using various predictive algorithms and a panel of 4 peptides were acquired. Their oligonucleotides were designed, synthesized and inserted into the multiple cloning site of plasmid pET-32c(+). The recombinant plasmids were transformed into E.coli BL21 and identified by endonuclease digestion and sequencing. The positive clones containing the recombinant plasmids could express specific fusion proteins (trx-epitope, MW≈20kDa) induced by IPTG. The fusion protein with 6×His could be coupled with NTA resin specifically, and purified by elusion of the column with buffer containing imidazole. The purified fusion proteins were incubated with splenocytes of C3H mice and then, the proliferation of splenocytes was determined by^3H-TdR incorporation assay. Results The recombinant plasmids were constructed successfully and the positive clones containing the recombinant plasmids expressed specific fusion proteins. Three of the purified fusion proteins (P4、P5、P6) could stimulate the lymphocyte proliferation. Conclusion Three T cell epitopes on Sj22.6 antigen were identified.
出处 《中国寄生虫学与寄生虫病杂志》 CAS CSCD 北大核心 2003年第6期345-348,共4页 Chinese Journal of Parasitology and Parasitic Diseases
基金 国家自然科学基金课题 (No .30 2 71 1 66)~~
关键词 日本血吸虫 22.6kDa抗原 T细胞表位 血吸虫病 鉴定 克隆 Schistosoma japonicum, 22.6 kDa antigen, T cell epitope, identification
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参考文献7

  • 1Bergquist R, Al-Sherbiny M, Barakat R, et al. Blueprint for schistosomiasis vaccine development[J]. Acta Trop,2002,82:183 - 92.
  • 2张桂筠,张兆松,陈淑贞,沈一平,吴海玮,苏川,王荣芝,吴观陵.日本血吸虫中国大陆株基因重组抗原Sj 22.6kDa的核苷酸序列分析[J].中国寄生虫学与寄生虫病杂志,1998,16(2):105-108. 被引量:7
  • 3苏川,马磊,王荣芝,胡雪梅,陈淑贞,邵莉君,吴海玮,沈蕾,张兆松,吴观陵.日本血吸虫22.6kDa重组蛋白对小鼠的免疫保护性研究[J].中国寄生虫学与寄生虫病杂志,1999,17(5):288-291. 被引量:14
  • 4Waine GJ,Mazzer DR,Brandt ER,et al.A dominant B-cell epitope on the 22 kDa tegumental membrane-associated antigen of Schitosoma japonicummaps to an EF-hand calcium binding domain[J].Parasite Immunol,1997,19:337-45.
  • 5Zhouj,Waine GJ,Zeng Q,etal.B-cell epitopes recognized by Chinese water buffaloes(Bos buffelus)onthe 22 kDategumental membrane-associated antigen(Sj-22)ofthe Asiatic blood fluke,Schistosomajaponicum [j].Vet Res,1999,30 :427-32.
  • 6胡雪梅,张兆松,吴海玮,苏川,季旻珺,王勇,陈淑贞,吴观陵.用噬菌体随机肽库鉴定日本血吸虫22.6kDa抗原分子的表位[J].中国寄生虫学与寄生虫病杂志,2002,20(3):164-167. 被引量:8
  • 7Schirle M, Weinschenk T, Stevanovic S. Combining computer algorithms with experimental approaches permits the rapid and accurate identification of T cell epitopes from defined antigens [ J ]. J Immunol Methods,2001, 257:1 - 16.

二级参考文献21

  • 1张桂筠,张兆松,陈淑贞,沈一平,吴海玮,苏川,王荣芝,吴观陵.日本血吸虫重组抗原基因的高效表达和特性鉴定[J].中国人兽共患病杂志,1996,12(5):30-33. 被引量:8
  • 2萨姆布鲁克 金冬雁(译).分子克隆实验指南(第二版)[M].北京:科学出版社,1993.19-21.
  • 3张桂筠,中国寄生虫学与寄生虫病杂志,1998年,16卷,6页
  • 4张桂筠,中国寄生虫学与寄生虫病杂志,1997年,15卷,326页
  • 5Cai Mengshen,Workshop on Molecular Biology of Schistosoma japomicum,1994年
  • 6鲁润龙,细胞生物学杂志,1991年,109页
  • 7赵元寿,细胞分子生物学,1990年,329页
  • 8苏川,中国寄生虫学与寄生虫病杂志,1999年,17卷,205页
  • 9张桂筠,中国寄生虫学与寄生虫病杂志,1998年,16卷,105页
  • 10张桂筠,中国寄生虫学与寄生虫病杂志,1998年,16卷,6页

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