摘要
为有效分离实验系统中同时存在于哺乳类细胞中的两种质粒 ,利用插入失活对真核细胞表达质粒的抗性进行改建 .以具有编码氨苄抗性基因的p REP9质粒和同时有编码氨苄和卡那抗性基因的p MAMneo质粒为起始模板得到无氨苄抗性而具有卡那抗性的真核细胞表达质粒 :p REP9- amp-和p MAMneo- amp-.前者可用于外源基因在细胞内的持续表达而后者对外源基因表达受地塞米松的诱导 .当细胞中同时转染有其他抗性的质粒时 ,可以通过抗生素抗性的不同而使两种质粒得到有效地分离和筛选 ,并能与 p Z1 89穿梭质粒配套应用于突变研究 .
In order to separate two kinds of plasmids existing in the same mammalian cell simultaneously in a certain experiment system, inserting inactivation was used to reconstruct eukaryotic cell expression plasmids and change their antibiotic resistance. Using pREP9 that contains ampicillin resistance and pMAMneo that contains ampicillin and kanamycin resistance as template, we got two new eukaryotic cell expression plasmids: pREP9 amp - and pMAMneo amp - which can be selected in E.coli by kanamycin instead of ampicillin. pREP9 amp - can be used for consistent expression while the latter one directs dexamethasone inducible expression. These vectors can be isolated from other plasmids through antibiotic resistant selection conveniently when the cells were transfected with two kinds of plasmids and compatible with the mutation detection system of shuttle plasmid pZ189.
出处
《中国药理学与毒理学杂志》
CSCD
北大核心
2000年第2期144-148,共5页
Chinese Journal of Pharmacology and Toxicology
基金
国家自然科学基金资助 !(39830 2 10 )&&
