Baranov V I, Ortlepp S A, Spirin A S
Insitute of Protein Research, Academy of Sciences of the USSR, Moscow Region.
Gene. 1989 Dec 14;84(2):463-6. doi: 10.1016/0378-1119(89)90521-0.
A cell-free system for preparative gene expression is described. It is composed of DNA-free Escherichia coli extract and added plasmid DNA; coupled transcription-translation proceeds with a continuous flow of the feeding solution containing nucleoside triphosphates and amino acids. The system works at a high constant rate for tens of hours. The yield of synthesised proteins after 20-50 h is hundreds of micrograms from 1 ml of the reaction mixture. Electrophoretic analysis of translation products confirms synthesis of proteins of the expected molecular mass.
描述了一种用于制备性基因表达的无细胞系统。它由不含DNA的大肠杆菌提取物和添加的质粒DNA组成;转录-翻译偶联过程通过含有核苷三磷酸和氨基酸的进料溶液的连续流动进行。该系统能以高恒定速率工作数十小时。在20至50小时后,从1毫升反应混合物中合成蛋白质的产量为数百微克。对翻译产物的电泳分析证实了预期分子量蛋白质的合成。