Characterisation and development of aspirin inducible biosensors in E. coli Nissle 1917 and SimCells
Characterisation and development of aspirin inducible biosensors in E. coli Nissle 1917 and SimCells
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大肠杆菌 Nissle 1917 和 SimCells 中阿司匹林诱导生物传感器的表征和开发
DOI：
10.1101/414169
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Chen J
中科院分区：
文献类型：
--
作者：
Chen J
利用从不动杆菌baylyiADP1克隆的Psal启动子和SalR调控系统，建立了一个简单的阿司匹林诱导系统。基因回路的大肠杆菌鉴定和新的SimCells(简单细胞)的诱导。在SalR蛋白INA的DNA结合域(DBD)和化学识别域(CRD)处发生突变。BaylyiADP1提示失活的Salrican与激活的SalRa竞争，占据Psal启动子的结合位置。在两种不同的设计中比较了Psal启动子的诱导。Coli：简单调节(SRS)和正自动调节系统(PAR)。当阿司匹林浓度在0.0 5~10μM范围内，两种调控系统均以剂量依赖的方式被诱导。弱的SalR表达显著提高了诱导强度，这与SalRi/SalRi竞争性结合的假说很好地一致。PAR系统提供了一个反馈回路，微调SAR的水平，显示出可诱导的强度。建立了一个基于SalRi/SalRi竞争性结合假说的数学模型，该模型不仅再现了观察到的实验结果，而且预测了一种新的基因电路设计的性能。阿司匹林诱导系统在益生菌E.coliNissle1917(ECN)和Frome生产的SimCells中也具有功能。ColiMC1000ΔMind。阿司匹林诱导的基因电路具有良好的特性和模块化，将成为环境和医疗应用中细菌治疗的有用生物砖。
A simple aspirin-inducible system has been developed by employing the Psalpromoter and SalR regulation system originally fromAcinetobacter baylyiADP1, which has been cloned intoE. colifor characterisation of gene circuits and induction of novel SimCells (simple cells). Mutagenesis at the DNA binding domain (DBD) and chemical recognition domain (CRD) of the SalR protein inA. baylyiADP1 suggests that inactive SalRican compete with activated SalRa, occupying the binding position of Psalpromoter. The induction of the Psalpromoter was compared in two different designs inE. coli: simple regulation (SRS) and positive autoregulated system (PAR). Both regulatory systems were induced in a dose-dependent manner in the presence of aspirin in the range of 0.05-10 μM. Over-expression of SalR in the SRS system reduces both baseline leakiness and inducible strength of Psalpromoter. A weak SalR expression significantly improve the inducible strength, which is in a good agreement of the proposed hypothesis of SalRi/SalRacompetitive binding. The PAR system provides a feedback loop that fine-tunes the level of SalR, displaying inducible strength. A mathematical model based on SalRi/SalRacompetitive binding hypothesis was developed, which not only reproduces the observed experimental results but also predict the performance of a new gene circuit design. The aspirin-inducible systems were also functional in probiotic strainE.coliNissle 1917 (EcN) and SimCells produced fromE. coliMC1000 ΔminD. The well-characterised and modularised aspirin-inducible gene circuits would be useful biobricks for bacterial therapy in environment and medical applications.
影响因子：
5.6
作者：
Mitrophanov AY;Hadley TJ;Groisman EA
通讯作者：
Groisman EA
DOI：
10.1073/pnas.1319470110
发表时间：
2013-12-10
影响因子：
11.1
作者：
Reyes, Alejandro;Wu, Meng;Gordon, Jeffrey I.
通讯作者：
Gordon, Jeffrey I.
阿司匹林治疗和预防心血管疾病的最新进展。
影响因子：
4.8
作者：
C. Hennekens
通讯作者：
C. Hennekens