Synergism in replication and translation of messenger RNA in a cell-free system.
Synergism in replication and translation of messenger RNA in a cell-free system.
复制标题
无细胞系统中信使 RNA 复制和翻译的协同作用。
DOI：
10.1073/pnas.90.20.9325
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发表时间：
1993
影响因子：
11.1
通讯作者：
A. Spirin
中科院分区：
文献类型：
--
作者：
I. Morozov;V. I. Ugarov;A. Chetverin;A. Spirin
Q β复制酶反应与大肠杆菌无细胞翻译系统的组合显著增强了重组RQ-DHFR RNA的复制，所述重组RQ-DHFR RNA由插入到RQ 135（-1）RNA中的二氢叶酸还原酶（DHFR）mRNA序列组成，RQ 135（-1）RNA是有效的天然存在的Q β复制酶模板。这种增强与先前描述的体内Q β噬菌体RNA复制的复制不对称性有关;正义（+）链的产生远远超过反义（-）链。这反过来又导致功能活性DHFR的合成增加。当DHFR mRNA或RQ 135（-1）RNA用作模板时，如果翻译系统不完整，或如果它被嘌呤霉素抑制，则未观察到这些效应。非病毒mRNA重组体的耦合复制-翻译可以作为研究病毒扩增的基本方面的有用模型，并且可以用于体外大规模蛋白质合成。
Combination of the Q beta replicase reaction with the Escherichia coli cell-free translation system markedly enhances replication of a recombinant RQ-DHFR RNA consisting of the dihydrofolate reductase (DHFR) mRNA sequence inserted into RQ135(-1) RNA, an efficient naturally occurring Q beta replicase template. The enhancement is associated with a replication asymmetry previously described for the replication of Q beta phage RNA in vivo; the sense (+)-strands are produced in large excess over the antisense (-)-strands. This, in turn, results in increased synthesis of the functionally active DHFR. These effects are not observed when DHFR mRNAs or RQ135(-1) RNAs are used as templates, if the translation system is not complete, or if it is inhibited by puromycin. The coupled replication-translation of nonviral mRNA recombinants can serve as a useful model for studying the fundamental aspects of virus amplification and can be implemented for large-scale protein synthesis in vitro.