密码子优化的轮状病毒VP6蛋白在烟草中的高效表达
发布时间：2018-10-24 09:20
【摘要】： 轮状病毒(Rotavirus,RV)是引起全世界婴幼儿和幼龄动物严重腹泻的最重要、最常见的病原。在发展中国家,每年因RV感染发生中度和严重腹泻以及死亡的儿童高达1887万人;在发达国家,因RV感染造成的经济损失是一个不容忽视的问题。据估计美国每年就此一项的医药损失高达2亿6千万美元。鉴于RV危害严重且无有效的治疗手段,世界卫生组织(WHO)将Rv疫苗列为最优先发展的疫苗项目之一。我们对于新型RV疫苗的研究仍在探索之中。 植物生物反应器是一种安全和廉价的生物系统。随着分子生物学的发展,特别是植物遗传工程的快速发展,人类可以通过自身的意愿设计和改造植物,高效的生产出廉价的人类生产和生活所需要的各种产品。这种植物反应器的概念,尤其是利用植物生产药用蛋白和工业化酶类的概念,以及植物直接作为口服疫苗的概念,渐渐受到人们的重视和慢慢被人们广泛的接受。 本实验尝试利用烟草作为生物反应器生产VP6蛋白。将密码子优化的VP6基因克隆到质粒中,构建植物表达载体pBI121-V6(U)。利用农杆菌介导法转化烟草,经卡那霉素(Kan)多重筛选,获得抗性植株,对抗性植株提取DNA进行PCR分析,表明VP6基因已经整合到烟草的基因组中。转基因植株的western blot显示pBI121-V6(U)的转基因烟草可以有效的表达VP6蛋白。实验结果表明,我们所构建的重组轮状病毒多表位具有一定的免疫原性。本研究为研制重组轮状病毒抗原表位亚单位疫苗、转基因植物口服疫苗提供一条新的途径和方法,具有一定的理论意义和应用前景。
[Abstract]:Rotavirus (Rotavirus,RV) is the most important and common cause of severe diarrhea in infants and young animals around the world. In developing countries, as many as 18.87 million children die each year from moderate and severe diarrhoea from RV infection, and economic losses from RV infection are a problem that cannot be ignored in developed countries. The United States is estimated to lose $260 million a year on this item. In view of the serious harm of RV and the lack of effective treatment, (WHO) of the World Health Organization (WHO) lists Rv vaccine as one of the most priority vaccine projects. We are still exploring new RV vaccines. Plant bioreactor is a safe and cheap biological system. With the development of molecular biology, especially the rapid development of plant genetic engineering, human beings can design and transform plants through their own will, and efficiently produce all kinds of products needed by cheap human production and life. The concept of this kind of plant reactor, especially the concept of using plants to produce medicinal proteins and industrializing enzymes, and the concept of plants as oral vaccines, have gradually been paid more and more attention to and accepted widely by people. In this experiment, tobacco was used as a bioreactor to produce VP6 protein. The codon optimized VP6 gene was cloned into the plasmid and the plant expression vector pBI121-V6 (U). Was constructed. Transgenic tobacco was transformed by Agrobacterium tumefaciens and kanamycin (Kan) was used to obtain resistant plants. DNA was extracted from resistant plants for PCR analysis. The results showed that VP6 gene had been integrated into the genome of tobacco. Western blot of transgenic plants showed that pBI121-V6 (U) transgenic tobacco could express VP6 protein effectively. The results showed that the recombinant rotavirus polyepitopes were immunogenicity. This study provides a new approach and method for the preparation of recombinant rotavirus epitope subunit vaccine and oral vaccine for transgenic plants, which has certain theoretical significance and application prospect.
【学位授予单位】：天津师范大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392;Q943.2
本文编号：2290974
[Abstract]:Rotavirus (Rotavirus,RV) is the most important and common cause of severe diarrhea in infants and young animals around the world. In developing countries, as many as 18.87 million children die each year from moderate and severe diarrhoea from RV infection, and economic losses from RV infection are a problem that cannot be ignored in developed countries. The United States is estimated to lose $260 million a year on this item. In view of the serious harm of RV and the lack of effective treatment, (WHO) of the World Health Organization (WHO) lists Rv vaccine as one of the most priority vaccine projects. We are still exploring new RV vaccines. Plant bioreactor is a safe and cheap biological system. With the development of molecular biology, especially the rapid development of plant genetic engineering, human beings can design and transform plants through their own will, and efficiently produce all kinds of products needed by cheap human production and life. The concept of this kind of plant reactor, especially the concept of using plants to produce medicinal proteins and industrializing enzymes, and the concept of plants as oral vaccines, have gradually been paid more and more attention to and accepted widely by people. In this experiment, tobacco was used as a bioreactor to produce VP6 protein. The codon optimized VP6 gene was cloned into the plasmid and the plant expression vector pBI121-V6 (U). Was constructed. Transgenic tobacco was transformed by Agrobacterium tumefaciens and kanamycin (Kan) was used to obtain resistant plants. DNA was extracted from resistant plants for PCR analysis. The results showed that VP6 gene had been integrated into the genome of tobacco. Western blot of transgenic plants showed that pBI121-V6 (U) transgenic tobacco could express VP6 protein effectively. The results showed that the recombinant rotavirus polyepitopes were immunogenicity. This study provides a new approach and method for the preparation of recombinant rotavirus epitope subunit vaccine and oral vaccine for transgenic plants, which has certain theoretical significance and application prospect.
【学位授予单位】：天津师范大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392;Q943.2
【引证文献】
相关硕士学位论文 前1条
1 邸萌亮;人脂联素基因（ADI）转化水稻的研究[D];扬州大学;2011年
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