日本血吸虫病诊断分子B细胞表位的预测与鉴定  

Prediction and identification of B cell epitopes of Schistosoma japonicum

作　　者：章辉[1] 司进[1] 朱荫昌[1] 赵松[1] 王晓婷[1] 殷旭仁[1] 曹利民[2] 华万全[1] 徐明[1] 梁幼生[1] 

机构地区：[1]江苏省血吸虫病防治研究所,江苏省寄生虫分子生物学重点实验室,江苏省寄生虫病学重点学科,无锡214064 [2]华中科技大学同济医学院

出　　处：《中国血吸虫病防治杂志》2006年第1期19-24,共6页Chinese Journal of Schistosomiasis Control

基　　金：江苏省卫生厅基金项目(X200318;X2111);江苏省预防医学科研项目(Y20020027)

摘　　要：目的 应用生物信息学技术对日本血吸虫病常用诊断分子进行B细胞表位的预测，筛选出具有潜在诊断价值的表位分子，采用基因工程方法制备目的表位的融合蛋白并进行抗原性鉴定。方法 将日本血吸虫膜蛋白22kDa（Sj22）、膜蛋白23kDa（Sj23）、信号蛋白14—3—3（Sj14—3—3）、谷胱甘肽S转移酶（Sj26）分子的全基因序列输入BioSun软件的工作区，经多参数比较筛选可能的B细胞表位，克隆、表达预测表位，用表达的融合蛋白作为抗原与血吸虫病人血清反应，以筛选和鉴定具有抗原性的表位。结果 经预测分析，Sj22的B细胞表位可能在56～62位和127～133位氨基酸区域，Sj23的B细胞表位可能在149～156位和160～167位氨基酸区域，Sj14—3—3的B细胞表位可能在118～225位和130～137位氨基酸区域，Sj26的B细胞表位可能在143～149位和191～197位氨基酸区域。克隆、表达并纯化这8个表位的融合蛋白，经SDS—PAGE和Western blot检测抗原性，筛选出分别来自于Sj22、Sj14-3-3、Sj26的3条具有较强抗原性的表位片段。结论 生物信息学技术结合分子生物学方法可用于筛选具有潜在诊断价值的表位。Objective To predict B cell epitopes in Sj22, Sj23, Sj14-3-3, Sj26 of Schistosoma japonicum with bioinformatics, and evaluate the antigenicity of these epitope proteins. Methods The complete DNA sequences of S. japonicum were predicted by BioSun system, the target B cell epitope genes were selected, cloned and expressed. The expressed fusion proteins were detected with the sera of schistosomiasis patients and health people for evaluation of their antigenicity, Results Eight B cell epitopes from four molecules of S. japonicum were predicted. The B cell epitopes of Sj22 probably located in 56-62 and 127 133 amino acids. The B cell epitopes of Sj23 probably located in 149-156 and 160-167 amino acids. The B cell epitopes of S14-3-3 probably located in 118-125 and 130-137 amino acids. The B cell epitopes of Sj26 probably located in 143-149 and 191- 197 amino acids, The predicted epitope genes were cloned into pET-32c plasmid and expressed. Three of eight expressed fusion proteins of epitopes were reacted with the sera of schistosomiasis patients but not with health people. Conclusion Three epitope antigens with potential diagnosis value are determined.

关 键 词：BioSun软件 预测 B细胞表位 克隆 表达 诊断 

分 类 号：R383.24[医药卫生—医学寄生虫学]