Heyde Keith C, Scott Felicia Y, Paek Sung-Ho, Zhang Ruihua, Ruder Warren C
Department of Mechanical Engineering, Carnegie Mellon University; Engineering Science and Mechanics Program, Virginia Polytechnic Institute and State University.
Department of Biological Systems Engineering, Virginia Polytechnic Institute and State University.
J Vis Exp. 2017 Mar 9(121):55300. doi: 10.3791/55300.
We have developed an abiotic-biotic interface that allows engineered cells to control the material properties of a functionalized surface. This system is made by creating two modules: a synthetically engineered strain of E. coli cells and a functionalized material interface. Within this paper, we detail a protocol for genetically engineering selected behaviors within a strain of E. coli using molecular cloning strategies. Once developed, this strain produces elevated levels of biotin when exposed to a chemical inducer. Additionally, we detail protocols for creating two different functionalized surfaces, each of which is able to respond to cell-synthesized biotin. Taken together, we present a methodology for creating a linked, abiotic-biotic system that allows engineered cells to control material composition and assembly on nonliving substrates.
我们开发了一种非生物-生物界面，使工程细胞能够控制功能化表面的材料特性。该系统由两个模块组成：一种经过合成工程改造的大肠杆菌菌株和一个功能化材料界面。在本文中，我们详细介绍了一种使用分子克隆策略对大肠杆菌菌株内选定行为进行基因工程改造的方案。一旦构建成功，该菌株在接触化学诱导剂时会产生更高水平的生物素。此外，我们详细介绍了创建两种不同功能化表面的方案，每种表面都能够对细胞合成的生物素做出反应。综上所述，我们提出了一种创建相连的非生物-生物系统的方法，该系统使工程细胞能够控制非生物基质上的材料组成和组装。
