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利用脂多胺技术通过重组f1噬菌体介导的哺乳动物细胞转染

Recombinant f1 phage-mediated transfection of mammalian cells using lipopolyamine technique.

作者信息

Yokoyama-Kobayashi M, Kato S

机构信息

Kanagawa Academy of Science and Technology, Japan.

出版信息

Anal Biochem. 1994 Nov 15;223(1):130-4. doi: 10.1006/abio.1994.1557.

Abstract

Recombinant f1 phages carrying a shuttle vector pKA1M for expression of blasticidin S deaminase were introduced into monkey COS7 cells by mixing with dioctadecylamidoglycylspermine (DOGS). Blasticidin S selection resulted in the detectable growth of resistant colonies within a week. The transfection efficiency depended on the amounts of the phage and DOGS, their ratio, and the time during which the cells were incubated with the phage/DOGS mixture. This method requires only several microliters of an Escherichia coli culture medium containing recombinant f1 phage particles and is applicable to various cell lines including mouse NIH/3T3, chinese hamster CHO-K1, and human HT-1080.

摘要

携带用于表达杀稻瘟菌素S脱氨酶的穿梭载体pKA1M的重组f1噬菌体,通过与二辛基酰胺基甘氨酰精胺(DOGS)混合被导入猴COS7细胞。经杀稻瘟菌素S筛选,一周内可检测到抗性菌落生长。转染效率取决于噬菌体和DOGS的量、它们的比例以及细胞与噬菌体/DOGS混合物孵育的时间。该方法仅需几微升含有重组f1噬菌体颗粒的大肠杆菌培养基,并且适用于包括小鼠NIH/3T3、中国仓鼠CHO-K1和人HT-1080在内的各种细胞系。

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