McDonald Nathan D, Rhea Katherine A, Berk Kimberly L, Zacharko Julie L, Miklos Aleksandr E
U.S. Army DEVCOM Chemical Biological Center, Aberdeen Proving Ground, Edgewood Area, Maryland 21010, United States.
ACS Synth Biol. 2021 Dec 17;10(12):3604-3607. doi: 10.1021/acssynbio.1c00505. Epub 2021 Dec 2.
Cellular lysates capable of transcription and translation have become valuable tools for prototyping genetic circuits, screening engineered functional parts, and producing biological components. Here we report that lysates derived from CO92 are functional and can utilize both the σ70 and the bacteriophage T7 promoter systems to produce green fluorescent protein (GFP). Because of the natural lifestyle of , lysates were produced from cultures grown at 21 °C, 26 °C, and 37 °C to mimic the infection cycle. Regardless of the promoter system the GFP production from 37 °C was the most productive and the 26 °C lysate was the least. When reactions are initiated with 5 nM of DNA, the GFP output of the 37 °C lysate is comparable with the productivity of other non- systems. The data we present demonstrate that, without genetic modification to enhance productivity, cell-free extracts from are functional and dependent on the temperature at which the bacterium was grown.
能够进行转录和翻译的细胞裂解物已成为用于构建遗传电路原型、筛选工程化功能部件以及生产生物组件的有价值工具。在此，我们报告从CO92获得的裂解物具有功能，并且可以利用σ70和噬菌体T7启动子系统来产生绿色荧光蛋白（GFP）。由于[具体细菌名称]的自然生活方式，裂解物是从在21°C、26°C和37°C下培养的培养物中产生的，以模拟感染周期。无论启动子系统如何，37°C培养物产生的GFP最多，26°C裂解物产生的最少。当反应以5 nM的DNA起始时，37°C裂解物的GFP产量与其他非[具体细菌名称]系统的生产力相当。我们展示的数据表明，无需进行基因改造来提高生产力，来自[具体细菌名称]的无细胞提取物具有功能，并且取决于细菌生长的温度。
