A novel T7 system utilizing mRNA coding for T7 RNA polymerase
A novel T7 system utilizing mRNA coding for T7 RNA polymerase
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DOI：
10.1016/s0006-291x(03)00076-7
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发表时间：
2003-02-21
影响因子：
3.1
通讯作者：
Mayumi, T
中科院分区：
文献类型：
--
作者：
Nakano, R;Nakagawa, T;Mayumi, T
T7 系统不需要将报告基因重新定位到细胞核以使其在细胞质中表达，而是依赖于 T7 RNA 聚合酶 (T7 RNAP) 酶和由 T7 启动子控制的报告基因 DNA 的共定位。在本研究中，我们开发了一种新的 T7 系统，与使用传统系统的系统相比，该系统的基因表达水平更高。将 β-珠蛋白基因的 5'- 和 3'- 非翻译区 (UTR) 插入报告基因可增强报告基因的表达，这可能是由于 mRNA 的稳定性和高效翻译所致。此外，用T7 RNAP mRNA（通过插入β珠蛋白5'-和3'-UTR序列以及帽和poly(A)尾结构进行修饰）转染细胞，而不是传统方法中使用的T7 RNAP蛋白，进一步增强了报告基因的表达。因此，这种使用 T7 RNAP mRNA 的新型 T7 系统对于细胞质中外源提供的 DNA 的有效基因表达可能非常有效。 (C) 2003 年爱思唯尔科学（美国）。版权所有。
The T7 system dose not require the relocation of a reporter gene to the nucleus for its gene expression in the cytoplasm, but relies on the co-localization of T7 RNA polymerase (T7 RNAP) enzyme and reporter gene DNA that is controlled by the T7 promoter. In the present study, we developed a new T7 system in that gene expression can occur at a higher level than those using conventional systems. Insertion of 5'- and 3'-untranslated regions (UTR) of beta-globin gene into a reporter gene enhanced the reporter gene expression, presumably due to the stability and efficient translation of the mRNA. Instead of the T7 RNAP protein used in conventional methods, moreover, transfection of cells with T7 RNAP mRNA, which has been modified by inserting beta-globin 5'- and 3'-UTR sequences as well as the cap and poly(A) tail structures, further enhanced the reporter gene expression. Thus, this novel T7 system using T7 RNAP mRNA may be powerful for the efficient gene expression of DNA exogenously provided in the cytoplasm. (C) 2003 Elsevier Science (USA). All right's reserved.