STUDY OF NUCLEOTIDE SEQUENCES ENCODING ANTIGENIC SITES

狂犬病病毒糖蛋白抗原位点的继续研究将 集中在单个氨基酸变化的位置， 糖蛋白的抗原变体，其区分参与 病毒中和抗体的结合。 比较核苷酸 序列分析将鉴定变异糖蛋白基因中的突变 其表达抗原性改变的糖蛋白。 这将是 通过在引物延伸实验中使用合成DNA引物来完成 从分离的病毒体RNA模板获得互补DNA转录物， 从抗原变异病毒。 推导的氨基酸取代 改变的核苷酸序列将与以下分组相关： 两株狂犬病病毒功能表位图谱的比较 糖蛋白 这项研究将扩展到比较抗原性 固定病毒与街头狂犬病病毒糖蛋白的决定簇。 氨基酸的地形关系与不同的 狂犬病病毒糖蛋白中的表位将基于（a）确定， 从核苷酸序列推断的残基变化的线性分布 分析，和（B）取代的残基诱导的效果， 克隆的糖蛋白基因的定点诱变。 的 作为抗原性的局部扰动的诱导变化的意义 将对结构进行评估。 构象和/或线性改变 衍生自突变基因的糖蛋白的结构将 其特征在于其结合单克隆抗体的能力。 最后，克隆的狂犬病病毒核蛋白的核苷酸序列 将确定双链互补DNA，并且序列 将研究编码抗原位点。 抗原变异 这反映在两种主要抗原的化学结构差异上 （核蛋白和糖蛋白）将定义有效的 疫苗预防和接触后治疗。

The continued study of rabies virus glycoprotein antigenic sites will concentrate on the location of single amino acid changes in the glycoprotein of antigenic variants which differentiate epitopes involved in the binding of virus-neutralizing antibodies. Comparative nuecleotide sequence analysis will identify mutations in variant glycoprotein genes which express antigenically altered glycoproteins. This will be accomplished by using synthetic DNA primers in primer extension experiments to obtain complementary DNA transcripts from virion RNA templates isolated from the antigenic variant viruses. Amino acid substitutions deduced from the altered nucleotide sequences will be correlated with the grouping of epitopes in the functional epitope maps of two fixed rabies virus strain glycoproteins. This study will be extended to compare antigenic determinants of fixed virus with street rabies virus glycoproteins. The topographic relationship of amino acids associated with the different epitopes in the rabies virus glycoprotein will be determined based on (a) the linear distribution of residue changes deduced from nucleotide sequence analysis, and (b) the effect of substituted residues induced by site-directed mutagenesis of the cloned glycoprotein gene. The significance of the induced change as a local perturbation of the antigenic structure will be evaluated. Conformational and/or linear alterations in the structure of the glycoprotein derived from the mutated gene will be characterized by its ability to bind monoclonal antibodies. Finally, the nucleotide sequence of a cloned rabies virus nucleoprotein double-stranded complementary DNA will be determined, and the sequences encoding antigenic sites will be investigated. Antigenic variations reflected by differences in chemical structure of the two major antigens (nucleoprotein and glycoprotein) will define limitations of effective vaccine prophylaxis and post-exposure treatment.