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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 effectively separate the two plasmids which exist in mammalian cells simultaneously in the experimental system, the eukaryotic expression plasmids were modified by insertional inactivation. The pPR9 plasmid with ampicillin resistance gene And kanamycin resistance gene p MAMneo plasmid as a starting template to obtain a non-ampicillin resistant kanamycin-resistant eukaryotic expression plasmid: p REP9-amp- and p MAMneo- amp-. The former can be used for foreign genes While the latter is induced by dexamethasone on the expression of foreign genes.When plasmids are transfected with other resistant plasmids at the same time, the two plasmids can be effectively separated by antibiotic resistance and Screening, and with pZ1 89 shuttle plasmid supporting the application of mutation research.