质粒拯救型T-DNA标签质粒的构建  

Construction of Rescue Plasmid with T-DNA Tag

作　　者：张金谌[1] 张学文[1] 戴雄泽 

机构地区：[1]湖南农业大学生物科学技术学院,湖南长沙410128 [2]湖南蔬菜工程技术重点实验室,湖南长沙410125

出　　处：《安徽农业科学》2007年第34期11016-11018,共3页Journal of Anhui Agricultural Sciences

基　　金：湖南蔬菜工程技术重点实验室开放基金资助

摘　　要：[目的]为了高效率地获得转基因植物的旁邻序列。[方法]以pUC18及pWM101为出发质粒,通过用限制性内切酶EcoRⅠ和HindⅢ双酶切质粒pUC18,得到含大肠杆菌复制起点及氨苄青霉素抗性基因的pUC18大片断,并将这段DNA整合到pWM101的T-DNA之中构建重组质粒。[结果]该质粒利用根癌农杆菌转化植物后,可利用潮霉素筛选转化细胞,拯救质粒则可转化大肠杆菌后,以氨苄青霉素进行筛选。[结论]该研究为以后的用T-DNA标签研究植物功能奠定了基础。[Objective] The aim of the research was to obtain the fire,king sequences in transgenic plants effectively. [Method] With pUC18 and pWM101 as initial plasmids, double enzme digestion was conducted on plasmid pUC18 with restriction endonueleases of EcoR Ⅰ and Hind Ⅲ to obtain large fragment of plastald pUC18 that contained replication origin of E. coli arid ampicillin resistance gene. Tbe DNA fragment was integrated into T-DNA of plasmid pWM101 to construct a recombinant plasmid. [ Result] Alter transforming plant by using Agrobacterium turnefaciens, the plasmid conld make use of hygromycin to screen the transformants. After transforming E. coli, the rescue plasmid could be screened by using ampicillin, [Conclusion] The research laid the foundation for studying plant functions by using T-DNA tags.

关 键 词：PUC18 pWM101 T-DNA标签 重组构建 

分 类 号：Q782[生物学—分子生物学]