【作者】 陈筑； 刘建国； 马欣荣； 张燕； 唐琳； 吴家媛； 张剑； 杨德琴；

【Author】 CHEN Zhu, LIU Jian-guo, MA Xin-rong, ZHANG Yan,JANG Lin,WU Jia-yuan, YANG De-qing. (Affiliated Stomatology Hospital, Zunyi Medical College. Zunyi 563003;Chengdu Biologic Research Institute,CAS, Chengdu 610041, Sichuan Province, China)

【机构】 遵义医学院附属口腔医院； 中国科学院成都生物研究所；

【摘要】 目的:构建变形链球菌葡糖基转移酶植物表达质粒p2355-gtfB、p2365-gtfB,为其转化植物研究奠定物质基础。方法:以真核表达质粒pcDNA3-gtfB为模板,通过PCR方法扩增含多个编码变形链球菌葡糖基转移酶抗原表位的目的基因gtfB,将回收产物经T-A克隆技术克降于中间载体pMD18-T,鉴定插入方向正确后,将目的基因从中间载体释放,再克隆至高效的植物表达载体p2355、p2365,用电转化法转化根癌农杆菌EHA105。结果:通过对重组质粒进行酶切验证和DNA序列测定分析,获得 3．7kb大小的目的基因gtfR,与预计大小相同,并定向插入植物表达载体,开放阅读框架正确。结论:成功构建了含多个葡糖基转移酶免疫显性抗原编码基因的植物表达质粒p2355-gtfB、p2365-gtfB,为下一步转化植物奠定了基础。更多还原

【Abstract】 OBJECTIVE: To construct plant effective expression plasmids p2355-gtfB, P2365-gtfB, which containing multiple catalytic sites and antigen epitopes of glucosyltransferase (GTF) of Streptococcus mutans , and prepare for the development of effective edible anti-caries vaccine. METHODS: Using plasmid pcDNA3-gtfB as template, obtained the gtfB by PCR amplification. Then the PCR products were inserted into plasmid inter- mediate vector pMD18-T, and then subcloned from the recombinant plasmid T-gtfB to plant expression vector p2355, p2365. Then p2355-gtfB and p2365 -gtfB were transformed into Agrobacterium tumefaciens EHA 105 respectively by electroporation. RESULTS: Through restriction enzyme mapping analysis and DNA sequencing , the construction of the plant expression recombinant plasmids p2355 -gtfB and p2365-gtfB with expected open reading frame was confirmed. CONCLUSION: The recombinant plasmids were constructed successfully, which encoded multiple catalytic sites and epitopes. This study will provide basis for further study on edible plants vaccine against dental caries.更多还原