产品详情
pCEP4-CAT哺乳表达质粒
基本信息
别名: pCEP4/CAT
启动子: CMV promoter
复制子: pUC ori
终止子: SV40 poly(A) signal
质粒分类: 哺乳细胞,蛋白过表达载体
质粒大小: 10891bp
原核抗性: 氨苄青霉素Amp
筛选标记: 潮霉素Hyg
克隆菌株: 大肠杆菌DH5α
培养条件: 37℃,有氧,LB
表达宿主: 哺乳细胞
诱导方式: 无须诱导,瞬时表达
5'测序引物: CMV-F:CGCAAATGGGCGGTAGGCGTG
3'测序引物: Sv40-polyA-R:GAAATTTGTGATGCTATTGC
质粒简介
pCEP4 is an episomal mammalian expression vector that uses thecytomegalovirus (CMV) immediate early enhancer/promoter for high level transcription of recombinant genes inserted into the multiple cloning site. The Epstein-Barr Virus replication origin (oriP) and nuclear antigen (encoded by the EBNA-1 gene) is carried by this plasmid to permit extrachromosomal replication in human, primate, and canine cells. pCEP4 also carries the hygromycin B resistance gene for stable selection in transfected cells.
pCEP4/CAT is provided as a positive control for the relative level of expression of recombinant proteins in a cell line of interest. It expresses the chloramphenicol acetyl transferase (CAT) protein from the CMV enhancer/promoter. Like its parent vector pCEP4, pCEP4/CAT contains the hygromycin B resistance gene for selection.
pCEP4是pCEP4/CAT的对照质粒,插入一个基因CAT.。
质粒图谱
pCEP4-CAT哺乳表达质粒使用说明:
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。
pCEP4-CAT哺乳表达质粒注意事项:
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。