Preparation of amino acid mixtures for cell-free expression systems
Preparation of amino acid mixtures for cell-free expression systems
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DOI：
10.2144/000114249
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发表时间：
2015-01-01
期刊：
影响因子：
2.7
通讯作者：
Noireaux, Vincent
中科院分区：
文献类型：
--
作者：
Caschera, Filippo;Noireaux, Vincent
在这里，我们提出了一个程序，用于制备氨基酸混合物，具有所需的组成和生理pH值，在高浓度的无细胞表达系统。高达2.1 mg/mL的活性蛋白质的合成在分批模式反应与所有大肠杆菌无细胞表达系统。与昂贵的商业试剂盒相比，我们的方法快速，易于执行，经济上具有优势，使其可用于高通量实验，非标准氨基酸的掺入和无细胞代谢工程。
Here we present a procedure for preparing amino acid mixtures-having both the desired composition and a physiological pH-at high concentrations for cell-free expression systems. Up to 2.1 mg/mL of active protein was synthesized in batch mode reactions with an all Escherichia coli cell-free expression system. Our method is fast, easy to execute, and economically advantageous compared to expensive commercial kits, making it useful for high-throughput experiments, incorporation of nonstandard amino acids, and cell-free metabolic engineering.