McCoy Kimberly, Douglas Trevor
Department of Chemistry, Indiana University, Bloomington, IN, USA.
Methods Mol Biol. 2018;1776:295-302. doi: 10.1007/978-1-4939-7808-3_20.
Protein cages are ubiquitous in nature and have been manipulated to encapsulate a range of nonnative cargos including organic, inorganic, and small molecules. Many protein cages are derived from virus capsids that have been rendered noninfectious through the preferential production and use of proteins that are solely involved in capsid assembly, but which do not encapsulate genetic material and therefore do not contribute to infectivity. Here, we describe the production of protein cargo(s) encapsulated inside of P22 virus-like particles (VLPs), derived from bacteriophage P22. This is achieved via genetic fusion of the cargo to a scaffolding protein, which becomes encapsulated in the P22 VLP during templated assembly of the protein cage.
蛋白质笼在自然界中无处不在,并且已被用于封装一系列非天然货物,包括有机、无机和小分子。许多蛋白质笼源自病毒衣壳,通过优先生产和使用仅参与衣壳组装但不封装遗传物质且因此不具有感染性的蛋白质,这些病毒衣壳已失去感染性。在这里,我们描述了包裹在源自噬菌体P22的P22病毒样颗粒(VLP)内部的蛋白质货物的生产。这是通过将货物与支架蛋白进行基因融合来实现的,该支架蛋白在蛋白质笼的模板组装过程中被封装在P22 VLP中。