产品详情
pPR3-N膜蛋白双杂交质粒
基本信息
启动子: CYC1 promoter
复制子: pUC ori,F1 ori,2μ ori
终止子: CYC1 terminator
质粒分类: 酵母细胞质粒;酵母杂交质粒;双杂交类质粒
质粒大小: 6200bp
质粒标签: N-NubG;N-HA
原核抗性: 氨苄青霉素Amp
筛选标记: TRP1
克隆菌株: DH5α等大肠杆菌
培养条件: 37℃,LB,有氧
表达宿主: 酵母菌
培养条件: 30℃,YPD,有氧
5'测序引物: CYC1-F:CTTTCCTTATACATTAGGACC
3'测序引物: CYC1-R:GGGACCTAGACTTCAGGTTG
备注: 高拷贝质粒
质粒简介
pPR3-N是一个用于膜蛋白酵母双杂交实验的质粒。The DUALmembrane system is intended for the detection and identification of interactions involving integral membrane proteins and membrane-associated proteins in yeast. The DUALmembrane pairwise interaction kit enables you to:
Investigate the interaction between a membrane protein (integral or membrane-associated) and a membrane protein or soluble protein
Map domains or amino acids which are critical for an interaction
Screen cDNA libraries using a membrane protein as a bait to find novel
interacting proteins (requires purchase of a separate cDNA library from Dualsystems)
质粒图谱
pPR3-N膜蛋白双杂交质粒使用说明:
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。
pPR3-N膜蛋白双杂交质粒注意事项:
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。