植物防龋疫苗融合基因表达载体中选择标记基因的去除  

Removal of the selective marker gene in the fusion gene expression vector of plant anti-caries vaccine

作　　者：郎遥玲 王倩 陈彬 白国辉 管晓燕 刘建国 Lang Yaoling;Wang Qian;Chen Bin;Bai Guohui;Guan Xiaoyan;Liu Jianguo(Guizhou Provincial Key Laboratory of Oral Disease Research in Colleges and Universities and Zunyi Key Laboratory of Oral Disease Research,School of Stomatology,Zunyi Medical University,Zunyi 563000,Guizhou Province,China;Guiyang Stomatological Hospital,Guiyang 550002,Guizhou Province,China)

机构地区：[1]贵州省普通高等学校口腔疾病研究特色重点实验室暨遵义市口腔疾病研究重点实验室,遵义医科大学口腔医学院,贵州省遵义市563000 [2]贵阳市口腔医院,贵州省贵阳市550002

出　　处：《中国组织工程研究》2024年第1期7-11,共5页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81260164),项目负责人:刘建国;贵州省委组织部贵州省第六批人才基地(RCJD2019-9),项目负责人:刘建国;遵义市首批市级人才基地(遵市科合社字[2017]20号),项目负责人:刘建国;贵州省教育厅青年科技人才成长项目(黔教合KY字[2022]276号),项目负责人:陈彬;贵州省卫生健康委科学技术基金项目(gzwkj2021-351),项目负责人:陈彬。

摘　　要：背景:出于对转基因植物的食用安全和生态安全的考虑,标记基因的存留是影响转基因植物的首要安全问题。目的:基于免疫防龋原理,课题组针对表面蛋白和葡糖基转移酶这2种致龋毒力因子,构建植物防龋疫苗融合基因表达载体pCAMBIA-E8-APB-DOCK8,为转基因植物疫苗的研发提供基础。方法:该研究运用DNA重组技术,通过DNA片段分离、连接、转化、克隆子检测、测序等系列步骤,将植物防龋疫苗融合基因表达载体pCAMBIA-E8-APB-DOCK8中的选择性标记基因Km和GUS去除。结果与结论:标记基因去除效率达99%,该研究为免疫防龋的转基因植物疫苗的安全生产奠定了良好的实验基础,也为其它植物疫苗的载体构建提供思路。BACKGROUND:In consideration of the food safety and ecological safety of transgenic plants,the retention of marker genes is the primary safety issue affecting transgenic plants.OBJECTIVE:Based on the principle of immune caries prevention,our research team successfully constructed the plant anti-caries vaccine fusion gene expression vector pCAMBIA-E8-APB-DOCK8 for these two caries causing virulence factors surface protein and glucosyltransferase,which provides a basis for the research and development of transgenic plant vaccine.METHODS:In this study,the selective marker genes Km and GUS in the plant caries vaccine fusion gene expression vector pCAMBIA-E8-APB-DOCK8 were removed by DNA recombination technology through a series of steps such as DNA fragment separation,connection,transformation,clone detection,and sequencing.RESULTS AND CONCLUSION:The efficiency of marker gene removal was 99%.This study has laid a good experimental foundation for the safe production of transgenic plant vaccine against dental caries,and also provided ideas for the construction of other plant vaccine vectors.

关 键 词：植物疫苗 免疫防龋 表达载体 标记基因 转基因安全 E8启动子 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R781.1