本文關(guān)鍵詞：帶有三聯(lián)密碼AAA的Kozak序列對(duì)品蹄疫DNA疫苗免疫效果的研究　出處：《延邊大學(xué)》2005年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： Kozak序列 口蹄疫 DNA疫苗 基因表達(dá)

【摘要】：目的:比較引入六種不同Kozak序列口蹄疫DNA疫苗的表達(dá)水平及免疫效果。方法:在口蹄疫DNA疫苗抗原編碼基因VP1起始位點(diǎn)引入6種不同的Kozak序列(KZ1～KZ6),插入真核表達(dá)載體pVAX1,制備口蹄疫DNA疫苗,對(duì)6-8周齡BLAB/C小鼠進(jìn)行免疫,用酶聯(lián)免疫吸附試驗(yàn)(enzyme_linked immunosorbent assay,ELISA),分析實(shí)驗(yàn)動(dòng)物外周血抗VP1的抗體效價(jià);同時(shí)插入帶有綠色熒光蛋白的融合表達(dá)載體pEGFP-N1,構(gòu)建6種體外表達(dá)質(zhì)粒,轉(zhuǎn)染HeLa細(xì)胞,進(jìn)行瞬時(shí)表達(dá)并分析體外表達(dá)水平。結(jié)果:引入Kozak序列KZ6的疫苗質(zhì)粒pKV6接種動(dòng)物后產(chǎn)生的免疫抗體水平高于其它組;相應(yīng)的體外表達(dá)質(zhì)粒pKG6轉(zhuǎn)染HeLa細(xì)胞后體外表達(dá)水平辦高于其它組。結(jié)論:引入Kozak序列KZ6(帶有三聯(lián)密碼AAA)的口蹄疫DNA疫苗,能最有效地增加疫苗的蛋白表達(dá)量,并達(dá)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.
【學(xué)位授予單位】：延邊大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2005
【分類號(hào)】：R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 吳海祥,鄭從義,屈三甫,郭荊哲,王燕麗;口蹄疫病毒誘導(dǎo)宿主細(xì)胞凋亡的研究[J];中國(guó)病毒學(xué);2001年02期

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