Modified S/MAR episomal vectors for stably expressing fluorescent protein-tagged transgenes with small cell-to-cell fluctuations

被引：8

作者：

Mizutani, Akiko ^{[1
,2
,3
]}

Kikkawa, Eri ^{[1
]}

Matsuno, Akira ^{[2
]}

Shigenari, Atsuko ^{[1
]}

Okinaga, Hiroko ^{[3
]}

Murakami, Mineko ^{[2
]}

Ishida, Hideyuki ^{[4
]}

Tanaka, Masafumi ^{[1
]}

Inoko, Hidetoshi ^{[1
]}

机构：

[1] Tokai Univ, Sch Med, Isehara, Kanagawa 2591193, Japan

[2] Teikyo Univ, Chiba Med Ctr, Dept Neurosurg, Ichihara, Chiba 2990111, Japan

[3] Teikyo Heisei Univ, Fac Hlth & Med Sci, Tokyo 1708445, Japan

[4] Tokai Univ, Dept Physiol, Sch Med, Isehara, Kanagawa 2591193, Japan

来源：

ANALYTICAL BIOCHEMISTRY | 2013年 / 443卷 / 01期

关键词：

Episomal vector; S/MAR; Transgene; Stable expression; SCAFFOLD/MATRIX-ATTACHED REGION; GENE-TRANSFER; TRANSCRIPTION; REPLICATION;

D O I：

10.1016/j.ab.2013.08.009

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：113 / 116

页数：4

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