Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
Nanotechnology. 2013 Jan 18;24(2):025605. doi: 10.1088/0957-4484/24/2/025605. Epub 2012 Dec 10.
The rod-shaped plant virus tobacco mosaic virus (TMV) is widely used as a nano-fabrication template, and chimeric peptide expression on its major coat protein has extended its potential applications. Here we describe a simple bacterial expression system for production and rapid purification of recombinant chimeric TMV coat protein carrying C-terminal peptide tags. These proteins do not bind TMV RNA or form disks at pH 7. However, they retain the ability to self-assemble into virus-like arrays at acidic pH. C-terminal peptide tags in such arrays are exposed on the protein surface, allowing interaction with target species. We have utilized a C-terminal His-tag to create virus coat protein-templated nano-rods able to bind gold nanoparticles uniformly. These can be transformed into gold nano-wires by deposition of additional gold atoms from solution, followed by thermal annealing. The resistivity of a typical annealed wire created by this approach is significantly less than values reported for other nano-wires made using different bio-templates. This expression construct is therefore a useful additional tool for the creation of chimeric TMV-like nano-rods for bio-templating.
杆状植物病毒烟草花叶病毒(TMV)被广泛用作纳米制造模板,其主要外壳蛋白上的嵌合肽表达扩展了其潜在的应用。在这里,我们描述了一种用于生产和快速纯化携带 C 末端肽标签的重组嵌合 TMV 外壳蛋白的简单细菌表达系统。这些蛋白质在 pH7 时不与 TMV RNA 结合或形成盘状结构,但它们保留了在酸性 pH 下自我组装成病毒样排列的能力。在这种排列中,C 末端肽标签暴露在蛋白质表面,允许与目标物种相互作用。我们利用 C 末端 His 标签创建了能够均匀结合金纳米粒子的病毒外壳蛋白模板纳米棒。这些可以通过从溶液中沉积额外的金原子来转化为金纳米线,然后进行热退火。通过这种方法制备的典型退火线的电阻率明显低于使用不同生物模板制备的其他纳米线的报道值。因此,该表达构建体是用于生物模板制备的类似 TMV 样纳米棒的嵌合构建体的有用附加工具。