STUDY OF NUCLEOTIDE SEQUENCES ENCODING ANTIGENIC SITES

狂犬病病毒糖蛋白抗原点的继续研究将 集中于单一氨基酸在体内的位置变化 识别相关表位的抗原变异体的糖蛋白 病毒中和抗体的结合。比较核苷酸 序列分析将识别变异糖蛋白基因的突变 表达抗原性改变的糖蛋白。这将是 利用合成DNA引物完成引物延伸实验 从分离的病毒粒子RNA模板中获得互补DNA转录本 从抗原性变异病毒。推导出的氨基酸取代 改变的核苷酸序列将与以下分组相关 两株狂犬病病毒固定株功能表位图中的表位 糖蛋白。这项研究将扩展到比较抗原性 街头狂犬病病毒糖蛋白固定病毒的决定因素。 与不同物种相关的氨基酸的拓扑关系 狂犬病病毒糖蛋白中的表位将根据(A)确定 由核苷酸序列推导出的残基变化的线性分布 分析，以及(B)由以下引起的取代残基的影响 克隆的糖蛋白基因的定点突变。这个 作为抗原局部扰动的诱导改变的意义 结构将被评估。的构象和/或线性变化 来自突变基因的糖蛋白的结构将是 以其结合单抗的能力为特征。 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，并将其序列 将对抗原位点进行编码调查。抗原性变异 反映在两种主要抗原的化学结构上的差异 (核蛋白和糖蛋白)将定义有效的限制 疫苗预防和暴露后治疗。

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.