Centre for Synthetic Biology and Innovation and Department of Bioengineering, Imperial College London, London SW7 2AZ, UK.
Nat Commun. 2011 Oct 18;2:508. doi: 10.1038/ncomms1516.
Modular and orthogonal genetic logic gates are essential for building robust biologically based digital devices to customize cell signalling in synthetic biology. Here we constructed an orthogonal AND gate in Escherichia coli using a novel hetero-regulation module from Pseudomonas syringae. The device comprises two co-activating genes hrpR and hrpS controlled by separate promoter inputs, and a σ(54)-dependent hrpL promoter driving the output. The hrpL promoter is activated only when both genes are expressed, generating digital-like AND integration behaviour. The AND gate is demonstrated to be modular by applying new regulated promoters to the inputs, and connecting the output to a NOT gate module to produce a combinatorial NAND gate. The circuits were assembled using a parts-based engineering approach of quantitative characterization, modelling, followed by construction and testing. The results show that new genetic logic devices can be engineered predictably from novel native orthogonal biological control elements using quantitatively in-context characterized parts.
模块化和正交的遗传逻辑门对于构建稳健的基于生物学的数字设备以定制合成生物学中的细胞信号非常重要。在这里，我们使用来自丁香假单胞菌的新型异型调节模块在大肠杆菌中构建了一个正交的与门。该设备由两个由单独启动子输入控制的共同激活基因 hrpR 和 hrpS 以及一个依赖于 σ(54)的 hrpL 启动子组成，该启动子驱动输出。只有当两个基因都表达时，hrpL 启动子才会被激活，从而产生数字式与门整合行为。通过将新的调控启动子应用于输入，并将输出连接到非门模块以产生组合的与非门，该与门被证明是模块化的。该电路是通过基于部件的工程方法进行定量表征、建模，然后进行构建和测试来组装的。结果表明，可以使用定量特征化的新型天然正交生物控制元件来预测性地设计新的遗传逻辑器件。
