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3500￥
获取电话pRI201-ON单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
pRI201-ON是一个单子叶植物发根农杆菌的双表达质粒。The Ri Plasmid pRI201 is designed for the transformation and expression of target genes from plant cells. This series of binary vectors for plant transformation retains the backbone of the pRI101 vectors, including an alcohol dehydrogenase (ADH) gene-derived 5' untranslated region (5'-UTR) downstream of the cauliflower mosaic virus (CaMV promoter)-derived 35S promoter. Additionally, these vectors have a heat shock protein (HSP) gene-derived terminator in place of the nopaline synthase (NOS) gene-derived terminator, allowing higher target gene expression compared with the pRI101 vector series. Multigene transformation with a single vector can be achieved by integrating an expression cassette containing another gene (promoter + enhancer + gene of interest + terminator) into the vector's second multiple cloning site (MCS2) located downstream of the HSP terminator.
The Ri Plasmid pRI201 binary vector series offers two types of vectors: pRI 201-AN DNA and pRI 201-ON DNA. pRI 201-AN DNA contains an Arabidopsis ADH-derived 5' UTR (AtADH 5'-UTR) and is suitable for dicotyledonous plant transformation. pRI 201-ON DNA contains a rice ADH-derived 5' UTR (OsADH 5'-UTR) and is suitable for monocotyledonous plant transformation. The pRI 201-AN DNA and pRI 201-ON DNA binary vectors for plant transformation have a mutant-type replication origin (Ri ori) from the Rhizobium rhizogenes Ri plasmid. These vectors also contain a replication origin (ColE1 ori) derived from the pUC plasmid; the ColE1 ori allows high copy number replication in E. coli. The vectors' multiple cloning site, located near the right border (RB) of T-DNA, allows stable target gene integration into the plant chromosome.
质粒图谱
pRI201-ON单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。
pRI201-ON单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。
