EXPRESSION OF ROTAVIRUS OUTER CAPSID PROTEIN VP4 BY A BACULOVIRUS RECOMBINANT

杆状病毒重组体表达轮状病毒外衣壳蛋白 VP4

• 批准号: 3809701
• 负责人: M GROZIGLIA
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3809701
• 关键词: Rotavirus; antibody neutralization test; antibody specificity; antigen antibody reaction; antiserum; biological polymorphism; complementary DNA; gene expression; genetic manipulation; genetic promoter element; genetic recombination; guinea pigs; immunization; insect virus; monoclonal antibody; protein sequence; serotyping; swine; transposon /insertion element; virus envelope; virus genetics; virus protein

Rotavirus outer capsid protein VP4 is a major independent protective antigen. A cDNA clone representing the VP4 gene of symptomatic human rotavirus strain KU (VP7 serotype 1) or DS-1 (VP7 serotype 2) or asymptomatic human rotavirus strain 1076 (VP7 serotype 2) was constructed and inserted into a baculovirus expression vector under the control of the polyhedrin promoter. The resulting recombinants expressed the appropriate authentic VP4 protein. Guinea pigs immunized with these VP4 proteins developed specific antibodies which neutralized infectivity of the rotavirus from which the immunizing VP4 was derived. These antisera were then used in neutralization tests to define the extent and distribution of VP4 antigenic polymorphism among human rotaviruses. Three distinct serotypes and one subtype of the VP4 outer capsid protein were identified among 17 human rotavirus strains that had previously been assigned to 5 distinct VP7 serotypes. For the most part, VP4 serotype segregated independently of VP7 serotype. Among the 10 rotavirus strains whose VP4 gene was previously sequenced, there was complete concordance between assignment of VP4 serotype by neutralization and classification according to VP4 amino acid homology. Thus, rotaviruses that exhibited a VP4 amino acid homology of 89% or greater belonged to the same VP4 serotype as determined by neutralization. Finally, evidence was obtained that the serotype-specific domain is located on the VP8 cleavage subunit of VP4.

轮状病毒外衣壳蛋白VP4是一种主要的独立保护蛋白 抗原。代表有症状人类 VP4 基因的 cDNA 克隆 轮状病毒株 KU（VP7 血清型 1）或 DS-1（VP7 血清型 2）或 构建无症状人轮状病毒株 1076（VP7 血清型 2） 并插入到杆状病毒表达载体的控制下 多角体启动子。所得重组体表达适当的 正宗的VP4蛋白。用这些 VP4 蛋白免疫的豚鼠 开发了特异性抗体，可以中和病毒的传染性 轮状病毒，免疫VP4源自该病毒。这些抗血清是 然后用于中和测试来定义的程度和分布 人类轮状病毒中的VP4抗原多态性。三个截然不同 鉴定出 VP4 外衣壳蛋白的血清型和一种亚型 在 17 种人类轮状病毒株中，之前被分配给 5 种 不同的 VP7 血清型。大多数情况下，VP4 血清型是分离的 与 VP7 血清型无关。 在此前已测序的10个VP4基因轮状病毒株中， VP4 血清型分配之间完全一致 根据VP4氨基酸同源性进行中和和分类。 因此，表现出 VP4 氨基酸同源性为 89% 或 通过中和测定，更大的属于相同的 VP4 血清型。 最后，获得证据表明血清型特异性结构域位于 位于 VP4 的 VP8 切割亚基上。