Jungle Express is a versatile repressor system for tight transcriptional control.
Jungle Express is a versatile repressor system for tight transcriptional control.
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Jungle Express是一种用于紧密转录控制的多功能阻遏系统。
DOI：
10.1038/s41467-018-05857-3
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发表时间：
2018-09-06
影响因子：
16.6
通讯作者：
Thelen MP
中科院分区：
文献类型：
--
作者：
Ruegg TL;Pereira JH;Chen JC;DeGiovanni A;Novichkov P;Mutalik VK;Tomaleri GP;Singer SW;Hillson NJ;Simmons BA;Adams PD;Thelen MP
严格调控的启动子对于许多生物应用是必不可少的，在这些应用中，强诱导性、可移植性和可扩展性是可取的。目前的系统往往不兼容大规模发酵由于高诱导剂成本和严格的介质要求。在这里，我们描述了“丛林快车”的自下而上的工程，一个表达系统，使有效的基因调控在不同的变形菌。该系统由eir引导，eir是一种多药物结合抑制因子，对其优化的操作符和阳离子染料具有高亲和力，可作为强大的诱导剂，成本可以忽略不计。在大肠杆菌中，工程启动子表现出最小的基础转录，并在1 μ M结晶紫的诱导下超过4个数量级，达到比当前最强细菌系统的表达水平更高的表达水平。此外，我们还提供了对EilR与其操作符和两种诱导剂的特定相互作用的分子见解。丛林快车的多功能性为严格控制和有效的基因表达开辟了道路，而不局限于宿主生物，底物或规模。具有强诱导性和可扩展性的严格调控启动子在生物学应用中是非常理想的。在这里，作者描述了“丛林快车”，一个基于eir抑制因子的广泛宿主系统，由阳离子染料激活。
Tightly regulated promoters are essential for numerous biological applications, where strong inducibility, portability, and scalability are desirable. Current systems are often incompatible with large-scale fermentations due to high inducer costs and strict media requirements. Here, we describe the bottom-up engineering of ‘Jungle Express’, an expression system that enables efficient gene regulation in diverse proteobacteria. This system is guided by EilR, a multidrug-binding repressor with high affinity to its optimized operator and cationic dyes that act as powerful inducers at negligible costs. In E. coli, the engineered promoters exhibit minimal basal transcription and are inducible over four orders of magnitude by 1 µM crystal violet, reaching expression levels exceeding those of the strongest current bacterial systems. Further, we provide molecular insights into specific interactions of EilR with its operator and with two inducers. The versatility of Jungle Express opens the way for tightly controlled and efficient gene expression that is not restricted to host organism, substrate, or scale. Tightly regulated promoters with strong inducibility and scalability are highly desirable for biological applications. Here the authors describe ‘Jungle Express’, a EilR repressor-based broad host system activated by cationic dyes.
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影响因子：
3.6
作者：
Fields AT;Navarrete CS;Zare AZ;Huang Z;Mostafavi M;Lewis JC;Rezaeihaghighi Y;Brezler BJ;Ray S;Rizzacasa AL;Barnett MJ;Long SR;Chen EJ;Chen JC
通讯作者：
Chen JC
影响因子：
6.4
作者：
Balzer S;Kucharova V;Megerle J;Lale R;Brautaset T;Valla S
通讯作者：
Valla S
影响因子：
10.5
作者：
EMORY, SA;BOUVET, P;BELASCO, JG
通讯作者：
BELASCO, JG
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
56.9
作者：
HENDRICKSON, WA
通讯作者：
HENDRICKSON, WA