Modified S/MAR episomal vectors for stably expressing fluorescent protein-tagged transgenes with small cell-to-cell fluctuations
Modified S/MAR episomal vectors for stably expressing fluorescent protein-tagged transgenes with small cell-to-cell fluctuations
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DOI：
10.1016/j.ab.2013.08.009
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发表时间：
2013-12-01
影响因子：
2.9
通讯作者：
Inoko, Hidetoshi
中科院分区：
文献类型：
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作者：
Mizutani, Akiko;Kikkawa, Eri;Inoko, Hidetoshi
我们修改和测试支架/基质附着区（S/MAR）附加型载体。新载体将有助于获得稳定表达荧光蛋白标记的转基因的细胞，其细胞间波动很小，大多在10倍以内。在载体中，相同的转录物指导附加型复制和转基因/抗生素标记物的表达，并且仅抗生素选择而没有任何其他额外步骤足以获得所需的稳定细胞，包括同时表达两种不同蛋白质的细胞。此外，这两个测试案例（AtT 20中人类生长激素的表达和HEK 293中四种蛋白激酶C亚型的表达）将被证明在可视化和分析涉及这些蛋白质的调控过程中是有用的。(C)2013 Elsevier Inc. All rights reserved.
We modified and tested scaffold/matrix attachment region (S/MAR) episomal vectors. The new vectors would be useful in obtaining cells stably expressing fluorescent protein-tagged transgenes with small, mostly within 10-fold cell-to-cell fluctuations. In the vectors, the same transcript directs episomal replication and expression of transgene/antibiotic marker, and only antibiotic selection without any other extra steps was sufficient to obtain desired stable cells, including those expressing two different proteins simultaneously. Furthermore, the two test cases (expression of human growth hormone in AtT20 and four protein kinase C isoforms in HEK293) would prove to be useful in visualizing and analyzing regulatory processes involving these proteins. (C) 2013 Elsevier Inc. All rights reserved.