带有三联密码AAA的Kozak序列对品蹄疫DNA疫苗免疫效果的研究

发布时间：2017-12-28 12:18

本文关键词：带有三联密码AAA的Kozak序列对品蹄疫DNA疫苗免疫效果的研究　出处：《延边大学》2005年硕士论文　论文类型：学位论文

更多相关文章： Kozak序列 口蹄疫 DNA疫苗 基因表达

【摘要】：目的:比较引入六种不同Kozak序列口蹄疫DNA疫苗的表达水平及免疫效果。方法:在口蹄疫DNA疫苗抗原编码基因VP1起始位点引入6种不同的Kozak序列(KZ1～KZ6),插入真核表达载体pVAX1,制备口蹄疫DNA疫苗,对6-8周龄BLAB/C小鼠进行免疫,用酶联免疫吸附试验(enzyme_linked immunosorbent assay,ELISA),分析实验动物外周血抗VP1的抗体效价;同时插入带有绿色荧光蛋白的融合表达载体pEGFP-N1,构建6种体外表达质粒,转染HeLa细胞,进行瞬时表达并分析体外表达水平。结果:引入Kozak序列KZ6的疫苗质粒pKV6接种动物后产生的免疫抗体水平高于其它组;相应的体外表达质粒pKG6转染HeLa细胞后体外表达水平办高于其它组。结论:引入Kozak序列KZ6(带有三联密码AAA)的口蹄疫DNA疫苗,能最有效地增加疫苗的蛋白表达量,并达到最佳的免疫效果。
[Abstract]:Objective: To compare the expression level and immune effect of six different Kozak sequences of foot-and-mouth disease DNA vaccine. Methods: the introduction of 6 different Kozak sequences in the DNA FMD vaccine antigen encoding gene VP1 codon (KZ1 ~ KZ6), inserted into the eukaryotic expression vector pVAX1, preparation of FMD DNA vaccine to immunize 6-8 BLAB/C mice by enzyme-linked immunosorbent assay (enzyme_linked immunosorbent, assay, ELISA) antibody the titer of anti serum VP1 analysis of experimental animal outside; while inserting fusion with green fluorescent protein expression vector pEGFP-N1, construct 6 kinds of in vitro expression plasmid transfected into HeLa cells for transient expression analysis and in vitro expression. Results: the antibody level of Vaccine Plasmid pKV6 with Kozak sequence KZ6 was higher than that of other groups after inoculation, and the expression level of corresponding plasmid pKG6 in vitro was higher than that of other groups. Conclusion: the introduction of Kozak sequence KZ6 (DNA with triple AAA) can effectively increase the protein expression of vaccine and achieve the best immune effect.
【学位授予单位】：延边大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392

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