Guo Yin Yin, Blocker Forrest, Xiao Feng, Guo Peixuan
Department of Pathobiology and Wildon School of Bioengineering, Purdue University, West Lafayette, IN 47907, USA.
J Nanosci Nanotechnol. 2005 Jun;5(6):856-63. doi: 10.1166/jnn.2005.143.
The bottom-up assembly of patterned arrays is an exciting and important area in current nanotechnology. Arrays can be engineered to serve as components in chips for a virtually inexhaustible list of applications ranging from disease diagnosis to ultrahigh-density data storage. In attempting to achieve this goal, a number of methods to facilitate array design and production have been developed. Cloning and expression of the gene coding for the connector of the bacterial virus phi29 DNA-packaging motor, overproduction of the gene products, and the in vitro construction of large-scale carpet-like arrays composed of connector are described in this report. The stability of the arrays under various conditions, including varied pH, temperature and ionic strength, was tested. The addition of packaging RNA (pRNA) into the array caused a dramatic shift in array structure, and resulted in the conversion of tetragonal arrays into larger decagonal structures comprised of both protein and RNA. RNase digestion confirmed that the conformational shift was caused by pRNA, and that RNA was present in the decagons. As has been demonstrated in biomotors, conformational shift of motor components can generate force for motor motion. The conformational shift reported here can be utilized as a potential force-generating mechanism for the construction of nanomachines. Three-dimensional computer models of the constructed arrays were also produced using a variety of connector building blocks with or without the N- or C-terminal sequence, which is absent from the current published crystal structures. Both the connector array and the decagon are ideal candidates to be used as templates to build patterned suprastructures in nanotechnology.
图案化阵列的自下而上组装是当前纳米技术中一个令人兴奋且重要的领域。可以设计阵列作为芯片中的组件，用于从疾病诊断到超高密度数据存储等几乎无穷无尽的一系列应用。为了实现这一目标，已经开发了许多促进阵列设计和生产的方法。本报告描述了编码细菌病毒phi29 DNA包装马达连接体的基因的克隆和表达、基因产物的过量生产以及由连接体组成的大规模地毯状阵列的体外构建。测试了阵列在各种条件下的稳定性，包括不同的pH值、温度和离子强度。向阵列中添加包装RNA（pRNA）会导致阵列结构发生显著变化，并导致四边形阵列转变为由蛋白质和RNA组成的更大的十边形结构。核糖核酸酶消化证实构象变化是由pRNA引起的，并且RNA存在于十边形结构中。正如在生物马达中所证明的那样，马达组件的构象变化可以产生驱动马达运动的力。这里报道的构象变化可作为构建纳米机器的潜在力产生机制。还使用了各种带有或不带有N端或C端序列的连接体构建模块制作了所构建阵列的三维计算机模型，而当前已发表的晶体结构中不存在这些序列。连接体阵列和十边形结构都是纳米技术中用作构建图案化超结构模板的理想候选者。
