产品介绍
pAdTrack-CMV腺病毒质粒
基本信息
启动子: CMV promoter
复制子: pUC ori
终止子: SV40 poly(A) signal
质粒大小: 9237bp
原核抗性: Kan
筛选标记: EGFP
克隆菌株: Stbl3
培养条件: 37℃,有氧,LB
诱导方式: 无须诱导,瞬时表达
质粒简介
PAdTrack-CMV transgenic adenovirus vector to cell expression vector with CMV promoter and EGFP gene expression of green fluorescent protein tracking results, and use common pAdeasy1 vector maximum can be inserted into the 5.9kb, the use of common pAdeasy2 carrier can insert the 8.6kb. The specific use of the method can refer to the pAdeasy carrier system.
Recombinant adenoviruses provide a versatile system for gene expression studies and therapeutic applications. We here report a strategy which simplifies the generation and production of such viruses. A recombinant adenoviral plasmid is generated with a minimum of enzymatic manipulations, employing homologous recombination in bacteria rather than in eucaryotic cells. Following transfections of such plasmids into a mammalian packaging cell line, viral production is conveniently followed with the aid of green fluorescent protein, encoded by a gene incorporated into the viral backbone. Homogeneous viruses can be obtained from this procedure without plaque purification.This system should expedite the process of generating and testing recombinant adenoviruses for a variety of purposes.
质粒图谱
pAdTrack-CMV腺病毒质粒使用说明:
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。
pAdTrack-CMV腺病毒质粒注意事项:
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。