工程菌DH_(5a)感受态细胞的制备及质粒pGEM的转化研究

Preparation of Engineering E Coli DH_(5a) Sensitive Cell and Study on the Plasmid pGEM Transformation

绿色荧光蛋白(green fluorescent protein,GFP)是源于海洋生物多管水母属(aequoia victoria)的一种发光蛋白,能够自身催化形成生色团并在蓝光激发下发出绿色荧光,能在活细胞内稳定表达,本试验首先从含有GFP的大肠杆菌中提取质粒,并利用胶回收试剂盒纯化质粒,采用氯化钙法将工程菌DH5a制成感受态细胞,然后将纯化后的质粒转入已制备好的感受态细胞中,使得工程菌DH5a转化成具有抗氨苄青霉素的大肠杆菌,并表达绿色荧光蛋白,接种于加氨苄青霉素的LB培养基上,筛选阳性克隆。

GFP to the optical bleaching, the oxidant, the reducing agent, the acid, the alkali and all that many chemical reagent has the greatly strengthened stability. First, from includes GFP in the backwoods coli to withdraw the material particle, and causes its purification using the rubber recycling reagent box. At the same time, makes using the calcium chloride law project fungus DHsa the feeling condition cell. Then will purify after the material particle to change over to already in the feeling condi- tion cell which the system will arrange, will cause project fungus DHsa to transform has the anti- ammonia animal pen penicillin the backwoods coli, and the vaccination in on the ammonification animal pen penicillin culture medium, willcause its growth.