摘要目的 确定汉滩病毒核衣壳蛋白(NP)B细胞表位的基因定位.方法 将DNase Ⅰ随机消化的汉滩病毒76/118株S基因片段与丝状噬菌体载体连接后转化MC1061菌,成功构建了汉滩病毒S基因-多肽片段噬菌体肽库.应用12株国产抗汉滩病毒单克隆抗体对表达汉滩病毒NP多肽片段的噬菌体文库进行了3～4轮淘筛(biopanning),并对阳性噬菌体克隆进行了扩增和测序.结果 确定了S基因产物--病毒核衣壳蛋白氨基端的一个B细胞线性抗原表位,该表位覆盖aa1～aa86的范围,相当于S基因bp37-294,其核心序列可能由aa15-66构成.结论 该线性表位能与多种抗汉滩病毒单克隆抗体发生交叉反应,是汉滩病毒NP的主要抗原表位.

abstractsObjective In order to construct the library of Hantaan virus S gene-peptide fragments on phages,screening the epitopes of nucleocapsid protein with monoclonal antibodies against Hantaviruses.Methods Hantaan virus S gene from recombinant plasmid pUC-HTNV-S was digested with DNase Ⅰ and ligated with filamentous phage vector that has been cut with Sfi Ⅰ.The ligation were transformed into E.coli MC1061 for constructing the phage library of Hantaan virus S gene-peptide fragments.After biopanning with some strains of monoclonal antibodies,the positive phage clones were picked out for sequencing.Results A linear epitope consisting of amino acid(aa)1-86 within nucleocapsid protein reading frame was determined and its core sequence consists of aa 15-66.Conclusion The linear epitope is a major and crossreactive antigenic epitope.The findings above are helpful for elucidating the relationship between molecular structure and function of Hantaviruses.