Silva-Rocha Rafael, de Lorenzo Víctor
Systems and Synthetic Biology Program, Centro Nacional de Biotecnología CSIC, Madrid, Spain.
Mol Biosyst. 2011 Aug;7(8):2389-96. doi: 10.1039/c1mb05094j. Epub 2011 May 17.
Whether biological or electronic, man-engineered computation is based on logic circuits assembled with binary gates that are interconnected to perform Boolean operations. We report here the rewiring of the SOS system of Escherichia in a fashion that makes the output of both the recA and lexA promoters to faithfully follow the pattern of a binary composite OR-NOT gate (ORN) in which the inputs are DNA damage (e.g. nalidixic acid addition) and IPTG as an exogenous signal. Unlike other non-natural gates whose implementation requires changes in genes and promoters of the genome of the host cells, this ORN was brought about by the sole addition of wild-type bacteria with a plasmid encoding a module for LacI(q)-dependent expression of lexA. Specifically, we demonstrate that the interplay between native, chromosomally-encoded components of the SOS system and the extra parts engineered in such a plasmid made the desired performance to happen without any modification of the core DNA-damage response network. It is thus possible to artificially interface autonomous cell networks with a predetermined logic by means of Boolean gates built with regulatory elements already functioning in the recipient organism.
无论是生物计算还是电子计算,人工设计的计算都是基于由二进制门组装而成的逻辑电路,这些二进制门相互连接以执行布尔运算。我们在此报告了对大肠杆菌SOS系统的重新布线,使得recA和lexA启动子的输出能够忠实地遵循二元复合或非门(ORN)的模式,其中输入为DNA损伤(如添加萘啶酸)和作为外源信号的IPTG。与其他非天然门不同,其实现需要改变宿主细胞基因组中的基因和启动子,而这种ORN是通过仅向野生型细菌添加携带编码用于LexA的LacI(q)依赖性表达模块的质粒实现的。具体而言,我们证明了SOS系统天然的、染色体编码的组分与在这种质粒中设计的额外部分之间的相互作用,使得在不对核心DNA损伤反应网络进行任何修改的情况下实现了所需的性能。因此,通过利用受体生物体中已经起作用的调控元件构建布尔门,有可能将自主细胞网络与预定逻辑进行人工连接。