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为了预测和筛选人禽流感H5N1毒株神经氨酸酶(NA)蛋白的B细胞表位,检测了人禽流感H5N1毒株NA基因核苷酸序列.采用Kyte-Doolittle的亲水性方案、Emini方案和Jameson-Wolf抗原指数方案,辅以对NA蛋白的二级结构中的柔性区域的分析,并用SPSS13.0进行聚类和相关分析,建立了一种统计学筛选程序,预测了H5N1病毒NA蛋白的B细胞表位,并对毒株GD-01-06的NA蛋白变异进行了分析.预测结果通过吴氏抗原指数和SWISS-MODEL软件进行评价.结果发现,预测并筛选最有可能的B细胞表位位于NA蛋白N端第120～137,81～84,408～415,273～282,429～432,356～368,46～55,146～155,341～350,198～209肽段,提示它们是B细胞表位的优势区段;含有糖基化位点NGT126～128的120～137肽段为首选的NA蛋白抗原表位;H5N1毒株NA蛋白第53位(Ⅰ)位点缺失,这有助于增加毒株GD-01-06的NA蛋白表面柔性,而且抗原表位扩大(VEP46～48→VEPISNTNFL46～55).采用SWISS-MODEL软件建立的N1蛋白模型有助于评价抗原表位预测.结果表明,用多参数预测以及用统计学筛选H5N1毒株NA蛋白的B细胞表位,可以为分子免疫学研究和药物筛选提供依据;广东GD-01-06毒株NA蛋白53位(Ⅰ)位点缺失可能增强该抗原表位的抗原性。
In order to predict and screen the B cell epitopes of neuraminidase (NA) protein of H5N1 strain of human bird flu, the NA gene nucleotide sequence of H5N1 strain of human bird flu was detected.Using Kyte-Doolittle’s hydrophilicity scheme, Emini Protocol and the Jameson-Wolf antigen index scheme, supplemented by analysis of the flexible regions in the secondary structure of the NA protein and clustering and correlation analysis using SPSS 13.0 established a statistical screening program to predict the H5N1 virus NA Protein B-cell epitopes and analyzed the NA protein variation of strain GD-01-06.The prediction results were evaluated by Wu’s antigen index and SWISS-MODEL software.The results showed that the most likely B The cytoepitopes were located at the N-terminal of N-terminal from the 120th-137th, the 1st to the 84th, the 4th, the 4th, the 4th, the 4th, the 4th, the 4th, The glycosylation sites NGT126 ~ 128 of the 120 ~ 137 peptide is the preferred NA protein epitope; H5N1 strain NA protein 53 (Ⅰ) site is missing, which helps to increase the strain GD-01-06 Of NA protein surface flexibility, and the expansion of antigenic epitopes (VEP46 ~ 48 → VEPISNTNFL46 ~ 55). The N1 protein model established by SWISS-MODEL software can be used to evaluate epitope prediction.The results show that using multiple parameters prediction and statistical screening of B cell epitopes of NA protein of H5N1 strain can be used for molecular immunology and drug screening Provide the basis; Guangdong Guangdong GD-01-06 strain NA protein 53 (Ⅰ) site may increase the antigen epitope antigenicity.