EXPRESSION/DISTRIBUTION OF CONSERVED/SEROTYPE-SPECIFIC EPITOPES ON ROTAVIRUS VP8

三个cDNA克隆，每个约300个核苷酸，包括 人轮状病毒KU株VP4蛋白的VP8区， 构建了重组表达载体pGEMEX-1，将其插入pGEMEX-1载体中，并在大肠杆菌中表达 大肠埃希菌。相应多肽的免疫印迹分析 代号A(氨基酸1至105)、B(氨基酸85至183)或C (氨基酸153到254)，用抗KU或Wa的血清(VP4)进行 1A型)、DS-1(VP41B型)或1076或M37(VP42型) 轮状病毒株。这些高免血清是在豚鼠身上制备的。 接种纯化病毒或表达的VP4蛋白。免疫印迹 分析表明，位于片段B的表位为VP4血清型 和亚型特异性，并提示该片段可能是 VP4的血清型和亚型特异性。相比之下，片段A和 C包含在不同人类轮状病毒之间保守的表位。 VP4血清型。观察到的高水平交叉反应 用表达的VP8蛋白的抗血清进行中和试验 来自对保守的A和C片段的抗体。

Three cDNA clones of approximately 300 nucleotides each and comprising the VP8 region of the VP4 protein of human rotavirus (HRV) strain KU, were constructed, inserted into the pGEMEX-1 plasmid and expressed in Escherichia coli. Immunoblot analysis of the corresponding peptides designated A (amino acid 1 to 105), B (amino acid 85 to 183), or C (amino acid 153 to 254), were performed using antisera to KU or Wa (VP4 serotype 1A), DS-1 (VP4 serotype 1B), or 1076 or M37 (VP4 serotype 2) rotavirus strains. These hyperimmune sera were prepared in guinea pigs inoculated with purified virus or expressed VP4 protein. The immunoblot analysis showed that epitopes located in fragment B were VP4 serotype and subtype specific and suggested that this fragment may be responsible for VP4 serotype and subtype specificity. In contrast, fragments A and C contained epitopes that were conserved among different human rotavirus VP4 serotypes. The high level of cross-reactivity observed in neutralization assay with antisera to the expressed VP8 protein may result from antibody to the conserved A and C fragments.