狂犬病毒糖蛋白和核蛋白主要免疫活性位点的质粒载体构建  

Construction of recombinant plasmid carrier including the principal immunocompetent domains of rabies virus glycoprotein and nucleoprotein

作　　者：杨裕华[1] 汪天虹[2] 赵世立[1] 解力[1] 

机构地区：[1]山东省卫生防疫站防疫科,山东济南250014 [2]山东大学微生物技术国家重点实验室,山东济南250100

出　　处：《疾病控制杂志》2003年第3期198-200,共3页Chinese Journal of Disease Control and Prevention

基　　金：山东省自然科学基金项目 (编号 Y96 C2 1 0 5 3)

摘　　要：目的　为进一步研究狂犬病毒糖蛋白和核蛋白免疫活性位点的免疫特性 ,构建了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狂犬病毒糖蛋白、核蛋白 ,以确定目的基因 ,用分子克隆的方法将目的基因片段插入质粒载体 ,最后测定重组质粒的核酸序列。结果　在大肠杆菌质粒 PU C1 8多克隆位点 Eco R 和 Bam H 之间 ,根据其阅读框架 ,插入了含有狂犬病毒糖蛋白主要免疫活性位点的目的基因合成片段 :G 免疫活性位点的第 1 4 9～ 1 6 0位氨基酸残基及 G 免疫活性位点的第 331～ 340位氨基酸残基所对应的碱基序列。在 KS质粒多克隆位点 Eco R 和 Bam H 间 ,插入了狂犬病毒核蛋白 -免疫活性位点第 374～ 383位氨基酸残基所对应的碱基序列。Objective To study the immunogenicity of glycoprotein and nucloprotien immunocompetent domains of rabies virus, Construct the recombinant plasmid carrier of Escherichia coli. Methods The target gene were determined by referene and using the program of protein hydropathy profile to analyse the glycoprotein and nucleoprotein of rabies virus, inserted the fragment of target gene into plasmid carrier by the method of molecular cloning, detected the nucleic acid sequence of recombinant plasmid. Results The synthesized fragment of target gene including the gene sequence of the principal immunocompetent domains of rabies virus glycoprotein: the regions of GⅡ base sequence corresponding to amino acid residue 149～160 and GⅢ with residue 331～340 were inserted into region between EcoRⅠ and BamHⅠ at polycloning sites of plasmid PUC 18 of Escherichia coli according to the reading frame, and inserted the fragment of immunocompetent domains of rabies virus nucleoprotein base sequence corresponding to amino acid residue 374 ～383 into region between EcoRⅠ and BamHⅠ at polycloning sites of plasmid KS. Conclusions To promote the basis for studying the immunogenicity of the domains of rabies virus and gene immunization.

关 键 词：狂犬病毒糖蛋白 免疫活性位点 质粒载体 免疫学 核蛋白类 

分 类 号：R373.9[医药卫生—病原生物学] R392[医药卫生—基础医学]