The assembly of protein machines in cells is precise, rapid, and coupled to protein synthesis with regulation in space and time. The assembly of natural and synthetic nanomachines could be similarly controlled by genetic programming outside the cell. Here, we present quasi-two-dimensional (2D) silicon compartments that enable programming of protein assembly lines by local synthesis from surface-immobilized DNA brushes. Using this platform, we studied the autonomous synthesis and assembly of a structural complex from a bacteriophage and a bacterial RNA-synthesizing machine. Local synthesis and surface capture of complexes provided high assembly yield and sensitive detection of spatially resolved assembly intermediates, with the 3D geometry of the compartment and the 2D pattern of brushes dictating the yield and mode of assembly steps. Localized synthesis of proteins in a single gene brush enhances their interactions, and displacement of their genes in separated brushes leads to step-by-step surface assembly. This methodology enables spatial regulation of protein synthesis, and deciphering, reconstruction and design of biological machine assembly lines.

蛋白质机器在细胞中的组装是精确，快速的，并且与时空调节的蛋白质合成相结合。天然和合成纳米机器的组装可以通过细胞外的遗传程序类似地控制。在这里，我们介绍了准二维（2D）硅隔室，该隔室能够通过从表面固定的DNA刷进行局部合成来对蛋白质装配线进行编程。使用该平台，我们研究了由噬菌体和细菌RNA合成机自动合成和组装结构复合物的过程。配合物的局部合成和表面捕获提供了高组装产率和对空间分辨的组装中间体的灵敏检测，其中隔室的3D几何形状和刷子的2D模式决定了组装步骤的产率和方式。单个基因画笔中蛋白质的局部合成增强了它们之间的相互作用，并且它们基因在单独的画笔中的置换导致逐步的表面组装。这种方法可以实现蛋白质合成的空间调节，以及生物机器装配线的解密，重建和设计。