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Orthogonal Regulatory Circuits for Escherichia coli Based on the γ-Butyrolactone System of Streptomyces coelicolor
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2018-03-06 00:00:00 , DOI: 10.1021/acssynbio.7b00425 Marc Biarnes-Carrera 1 , Chang-Kwon Lee 2 , Takuya Nihira 2, 3 , Rainer Breitling 1 , Eriko Takano 1
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2018-03-06 00:00:00 , DOI: 10.1021/acssynbio.7b00425 Marc Biarnes-Carrera 1 , Chang-Kwon Lee 2 , Takuya Nihira 2, 3 , Rainer Breitling 1 , Eriko Takano 1
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Chemically inducible transcription factors are widely used to control gene expression of synthetic devices. The bacterial quorum sensing system is a popular tool to achieve such control. However, different quorum sensing systems have been found to cross-talk, both between themselves and with the hosts of these devices, and they are leaky by nature. Here we evaluate the potential use of the γ-butyrolactone system from Streptomyces coelicolor A3(2) M145 as a complementary regulatory circuit. First, two additional genes responsible for the biosynthesis of γ-butyrolactones were identified in S. coelicolor M145 and then expressed in E. coli BL21 under various experimental conditions. Second, the γ-butyrolactone receptor ScbR was optimized for expression in E. coli BL21. Finally, signal and promoter crosstalk between the γ-butyrolactone system from S. coelicolor and quorum sensing systems from Vibrio fischeri and Pseudomonas aeruginosa was evaluated. The results show that the γ-butyrolactone system does not crosstalk with the quorum sensing systems and can be used to generate orthogonal synthetic circuits.
中文翻译：
基于天蓝色链霉菌γ-丁内酯系统的大肠杆菌正交调控电路
化学诱导转录因子广泛用于控制合成装置的基因表达。细菌群体感应系统是实现这种控制的流行工具。然而，已经发现不同的群体感应系统在它们自身之间以及与这些设备的主机之间存在串扰，并且它们本质上是泄漏的。在这里，我们评估来自天蓝色链霉菌A3(2) M145的 γ-丁内酯系统作为补充调节电路的潜在用途。首先，在天蓝色 S.coelicolor M145中鉴定了另外两个负责 γ-丁内酯生物合成的基因，然后在各种实验条件下在大肠杆菌BL21 中表达。其次，γ-丁内酯受体 ScbR 针对在大肠杆菌BL21。最后，评估了来自天蓝色链球菌的 γ-丁内酯系统与来自费氏弧菌和铜绿假单胞菌的群体感应系统之间的信号和启动子串扰。结果表明，γ-丁内酯系统不会与群体感应系统发生串扰，可用于生成正交合成电路。
更新日期：2018-03-06
中文翻译：
基于天蓝色链霉菌γ-丁内酯系统的大肠杆菌正交调控电路
化学诱导转录因子广泛用于控制合成装置的基因表达。细菌群体感应系统是实现这种控制的流行工具。然而，已经发现不同的群体感应系统在它们自身之间以及与这些设备的主机之间存在串扰，并且它们本质上是泄漏的。在这里，我们评估来自天蓝色链霉菌A3(2) M145的 γ-丁内酯系统作为补充调节电路的潜在用途。首先，在天蓝色 S.coelicolor M145中鉴定了另外两个负责 γ-丁内酯生物合成的基因，然后在各种实验条件下在大肠杆菌BL21 中表达。其次，γ-丁内酯受体 ScbR 针对在大肠杆菌BL21。最后，评估了来自天蓝色链球菌的 γ-丁内酯系统与来自费氏弧菌和铜绿假单胞菌的群体感应系统之间的信号和启动子串扰。结果表明，γ-丁内酯系统不会与群体感应系统发生串扰，可用于生成正交合成电路。
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