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从小鼠肥大细胞瘤P815中筛选高转染性变体。

Selection of highly transfectable variant from mouse mastocytoma P815.

作者信息

Van Pel A, De Plaen E, Boon T

出版信息

Somat Cell Mol Genet. 1985 Sep;11(5):467-75. doi: 10.1007/BF01534840.

Abstract

A tk- cell line derived from mouse mastocytoma P815 was transfected with a plasmid carrying a thymidine kinase gene. The tk+ cells were obtained at a frequency of 10(-6). From some of these tk+ transfectants it was possible to select tk- cells with BrdU so that these cells could be submitted again to transfection with the thymidine kinase gene. By repeating cycles of transfection and tk+ selection followed by reverse selection with BrdU, it was possible to obtain a stable variant having a 100-fold increase in transfection efficiency. This HTR (high transfection) variant shows a high efficiency of transfection (10(-4)) for the neomycin-resistance gene as well as for the thymidine kinase gene.

摘要

从小鼠肥大细胞瘤P815衍生的tk-细胞系用携带胸苷激酶基因的质粒进行转染。以10(-6)的频率获得了tk+细胞。从其中一些tk+转染子中,有可能用溴脱氧尿苷(BrdU)选择tk-细胞,以便这些细胞能够再次用胸苷激酶基因进行转染。通过重复转染和tk+选择的循环,随后用BrdU进行反向选择,有可能获得转染效率提高100倍的稳定变体。这种HTR(高转染)变体对新霉素抗性基因以及胸苷激酶基因显示出高转染效率(10(-4))。

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