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目的 :为进一步研究狂犬病毒糖蛋白 (G蛋白 )免疫活性位点的免疫特性 ,构建其大肠杆菌重组质粒载体。方法 :参考有关文献及用蛋白质亲疏水性分析程序分析 ,确定目的基因片段 ,并用分子克隆的方法将目的基因片段插入载体质粒 ,最后测定其核酸序列。结果 :在大肠杆菌质粒 PU C18的多克隆位点 Eco R 和 Bam H 之间 ,根据其阅读框架 ,插入了人工合成的目的基因片段 ,其含有狂犬病毒 G蛋白免疫活性位点 G 表位第 149～ 16 0位氨基酸残基及 G 表位第 331～ 340位氨基酸残基所对应的核酸序列。结论 :本研究构建了狂犬病毒 G蛋白两个主要免疫活性位点的重组质粒载体 ,为深入研究其免疫活性位点的免疫特性及基因免疫打下基础。
OBJECTIVE: To further study the immunological characteristics of rabies virus glycoprotein (G protein) immunologically active site, construct recombinant plasmid vector of E.coli. Methods: With reference to the literature and the analysis of protein hydropathicity and hydrophobicity, the target gene fragment was identified. The target gene fragment was inserted into vector plasmid by molecular cloning method, and the nucleic acid sequence was determined. Results: According to its reading frame, EcoR and Bam H were inserted into EcoR and Bam H of Escherichia coli plasmid PU C18. Synthetic gene fragments were inserted, which contained the rabies G protein immunologically active G-site, 149 ~160 amino acid residues and G epitopes 331 ~ 340 amino acid residues corresponding to the nucleic acid sequence. Conclusion: In this study, we constructed a recombinant plasmid vector containing two major immunoreactive sites of rabies virus G protein, which lays the foundation for further study on the immunological characteristics and gene immunization of rabies virus G protein.