Gorochowski Thomas E, van den Berg Eric, Kerkman Richard, Roubos Johannes A, Bovenberg Roel A L
DSM Biotechnology Center, P.O. Box 1, 2600 MA Delft, The Netherlands.
ACS Synth Biol. 2014 Mar 21;3(3):129-39. doi: 10.1021/sb4001245. Epub 2013 Dec 10.
Synthetic biology has developed numerous parts for the precise control of protein expression. However, relatively little is known about the burden these place on a host, or their reliability under varying environmental conditions. To address this, we made use of synthetic transcriptional and translational elements to create a combinatorial library of constructs that modulated expression strength of a green fluorescent protein. Combining this library with a microbioreactor platform, we were able to perform a detailed large-scale assessment of transient expression and growth characteristics of two Escherichia coli strains across several temperatures. This revealed significant differences in the robustness of both strains to differing types of protein expression, and a complex response of transcriptional and translational elements to differing temperatures. This study supports the development of reliable synthetic biological systems capable of working across different hosts and environmental contexts. Plasmids developed during this work have been made publicly available to act as a reference set for future research.
合成生物学已经开发出许多用于精确控制蛋白质表达的元件。然而，对于这些元件给宿主带来的负担，或者它们在不同环境条件下的可靠性，我们了解得相对较少。为了解决这个问题，我们利用合成转录和翻译元件创建了一个构建体组合文库，该文库可调节绿色荧光蛋白的表达强度。将这个文库与微生物反应器平台相结合，我们能够对两种大肠杆菌菌株在几个温度下的瞬时表达和生长特性进行详细的大规模评估。这揭示了两种菌株对不同类型蛋白质表达的稳健性存在显著差异，以及转录和翻译元件对不同温度的复杂反应。这项研究支持了能够在不同宿主和环境背景下工作的可靠合成生物学系统的开发。在这项工作中开发的质粒已公开提供，作为未来研究的参考集。
