产品介绍
pRI101-AN双子叶植物发根农杆菌质粒
基本信息
启动子: 35S
原核抗性: 卡那霉素Kan
筛选标记: 遗传霉素G418
克隆菌株: 大肠杆菌HB101
培养条件: 37℃
质粒简介
pRI101-AN载体是一种在双子叶植物细胞中表达外源基因的载体。利用发根土壤杆菌介导。pRI101 DNA vectors are binary plant transformation vectors intended for foreign gene expression in plant cells. These agrobacterium-mediated plant transformation vectors carry the 35S promoter of caμliflower mosaic virus (CaMV) and the 5’ non-coding region (5’-UTR) of the alcohol dehydrogenase (ADH) gene. The 5’-UTR of ADH functions as a translation enhancer in plants. There are two types of pRI101 DNA vectors, the pRI101-AN DNA vector, which carries the 5’-UTR of Arabidopsis ADH (AtADH 5’-UTR), and the pRI101 ON DNA vector, which carries the 5’-UTR of rice ADH (OsADH 5’-UTR). The pRI101-AN DNA vector is for dicotyledonous plants such as tobacco or Arabidopsis while the pRI101-ON DNA vector is for monocotyledonous plants such as rice.Although the pRI101 DNA vectors are shuttle vectors and replicate autonomously in E. coli and Rhizobium (Agrobacterium), they are high copy number plasmids because they contain the same replication origin as pUC-type plasmids (ColE1 ori). These vectors are also stably maintained in Rhizobium (Agrobacterium) containing the mutant-type replication origin Ri (Ri-ori). The pRI101 DNA vectors are capable of stably integrating target genes into plant chromosomes because the vector cloning sites are located closer to the Right Border (RB) of T-DNA than the selection marker (NPT II) of the plant, so the target gene is not deleted.
质粒图谱
pRI101-AN双子叶植物发根农杆菌质粒使用说明:
1、收到质粒干粉后请先5000rpm离心1min,再加入20μl无菌水溶解质粒,室温放置1min;
2、从-80℃冰箱中取出相应的感受态,置于冰盒上解冻,并做好标记;
3、取2μl质粒加至100μl感受态中,冰浴30min;
4、42℃热激90s,再冰浴2min;
5、加入900μl无抗的LB液体培养基,180rpm震荡培养45min;
6、6000rpm离心5min,仅留100ul上清混匀菌体沉淀;
7、混匀后的菌液加至对应抗性的LB平板上,倒入适量玻璃珠,涂匀液体;
8、将平板正向培养1h,再倒置培养12h~16h;
9、挑取单克隆菌落至对应抗性的LB液体培养基中,震荡培养12h~16h,根据实验需要提取质粒。
pRI101-AN双子叶植物发根农杆菌质粒注意事项:
1、如果您收到的是甘油菌种,请先四区划线,挑取单克隆培养。
2、如果第二天转化平板长的过多,请将质粒按比例稀释后再转化。