An easy cell-free protein synthesis system dependent on the addition of crude Escherichia coli tRNA.
An easy cell-free protein synthesis system dependent on the addition of crude Escherichia coli tRNA.
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一种简单的无细胞蛋白质合成系统，依赖于添加粗制大肠杆菌 tRNA。
DOI：
10.1093/oxfordjournals.jbchem.a022581
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发表时间：
2000
影响因子：
2.7
通讯作者：
H. Takaku
中科院分区：
文献类型：
--
作者：
T. Kanda;K. Takai;S. Yokoyama;H. Takaku
大肠杆菌的蛋白质合成S30提取物含有tRNA，这限制了其在无细胞蛋白质合成中的应用。在这里，我们表明，通过固定化 RNase A 树脂处理，至少可以从标准 S30 提取物中去除 Arg 和 Ser 受体活性。这种经过 RNase 处理的提取物不具有蛋白质合成活性，但当与粗制大肠杆菌 tRNA 和少量人胎盘 RNase 抑制剂一起提供时，会恢复蛋白质合成活性。蛋白质合成依赖于在 RNase 抑制剂存在下添加 tRNA。用该系统合成了氯霉素乙酰转移酶，并发现该酶具有活性。
The protein-synthesizing S30 extract of Escherichia coli contains tRNA, which limits its applications in cell-free protein synthesis. Here, we show that at least Arg- and Ser-acceptor activities can be removed from a standard S30 extract by treatment with an immobilized RNase A resin. This RNase-treated extract exhibits no protein synthesis activity, but regains it when supplied with crude E. coli tRNA and a small amount of human placental RNase inhibitor. The protein synthesis is dependent on the addition of tRNA in the presence of the RNase inhibitor. Chloramphenicol acetyltransferase was synthesized with this system and found to be active.