Cell-free translation reconstituted with purified components
Cell-free translation reconstituted with purified components
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DOI：
10.1038/90802
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发表时间：
2001-08-01
影响因子：
46.9
通讯作者：
Ueda, T
中科院分区：
文献类型：
--
作者：
Shimizu, Y;Inoue, A;Ueda, T
我们开发了一种由纯化到均一的重组标签蛋白因子重组的蛋白质合成系统。该系统可以在不需要任何辅助设备的情况下，以分批方式生产蛋白质，速度约为160微克/毫升/小时。亲和层析法可在1h内对蛋白产物进行纯化，去除标记的蛋白因子。此外，省略释放因子允许使用抑制子转移RNA(TRNA)有效地掺入非天然氨基酸。
We have developed a protein-synthesizing system reconstituted from recombinant tagged protein factors purified to homogeneity. The system was able to produce protein at a rate of about 160 mug/ml/h in a batch mode without the need for any supplementary apparatus. The protein products were easily purified within 1 h using affinity chromatography to remove the tagged protein factors. Moreover, omission of a release factor allowed efficient incorporation of an unnatural amino acid using suppressor transfer RNA (tRNA).