Basic Medical Science and Molecular Medicine, Tokai University School of Medicine, Isehara, Kanagawa 259-1193, Japan; Department of Neurosurgery, Teikyo University Chiba Medical Center, Ichihara, Chiba 299-0111, Japan; Faculty of Health and Medical Science, Teikyo Heisei University, Higashi-Ikebukuro, Tokyo 170-8445, Japan.
Anal Biochem. 2013 Dec 1;443(1):113-6. doi: 10.1016/j.ab.2013.08.009. Epub 2013 Aug 19.
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.
我们修改并测试了支架/基质附着区（S/MAR）附加体载体。这些新载体对于获得稳定表达荧光蛋白标记的转基因细胞非常有用，其转基因/抗生素标记的拷贝数波动较小，大多数在 10 倍以内。在载体中，相同的转录本指导附加体复制和转基因/抗生素标记的表达，并且仅通过抗生素选择而无需任何其他额外步骤即可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（人生长激素在 AtT20 中的表达和四种蛋白激酶 C 同工型在 HEK293 中的表达）将被证明对于可视化和分析涉及这些蛋白质的调控过程非常有用。
