United States Department of Agriculture, Agricultural Research Service, Plant Sciences Institute, Molecular Plant Pathology Laboratory, Beltsville, MD 20705, USA.
J Virol Methods. 2011 Dec;178(1-2):209-15. doi: 10.1016/j.jviromet.2011.09.013. Epub 2011 Sep 22.
Maize rayado fino virus (MRFV) virus-like-particles (VLPs) produced in tobacco plants were examined for their ability to serve as a novel platform to which a variety of peptides can be covalently displayed when expressed through a Potato virus X (PVX)-based vector. To provide an anchor for chemical modifications, three Cys-MRFV-VLPs mutants were created by substituting several of the amino acids present on the shell of the wild-type MRFV-VLPs with cysteine residues. The mutant designated Cys 2-VLPs exhibited, under native conditions, cysteine thiol reactivity in bioconjugation reactions with a fluorescent dye. In addition, this Cys 2-VLPs was cross-linked by NHS-PEG4-Maleimide to 17 (F) and 8 (HN) amino acid long peptides, corresponding to neutralizing epitopes of Newcastle disease virus (NDV). The resulting Cys 2-VLPs-F and Cys 2-VLPs-HN were recognized in Western blots by antibodies to MRFV as well as to F and HN. The results demonstrated that plant-produced MRFV-VLPs have the ability to function as a novel platform for the multivalent display of surface ligands.
在烟草植物中产生的玉米线条病毒（MRFV）病毒样颗粒（VLPs）被检测其作为一种新型平台的能力，当通过基于马铃薯病毒 X（PVX）的载体表达时，各种肽可以通过共价方式展示在该平台上。为了提供化学修饰的锚点，通过将存在于野生型 MRFV-VLPs 外壳上的几个氨基酸替换为半胱氨酸残基，创建了三个 Cys-MRFV-VLPs 突变体。在天然条件下，指定为 Cys 2-VLPs 的突变体在与荧光染料的生物缀合反应中表现出半胱氨酸硫醇反应性。此外，该 Cys 2-VLPs 通过 NHS-PEG4-Maleimide 与 17（F）和 8（HN）个氨基酸长的肽交联，这些肽对应于新城疫病毒（NDV）的中和表位。用针对 MRFV 的抗体以及针对 F 和 HN 的抗体在 Western blot 中识别得到的 Cys 2-VLPs-F 和 Cys 2-VLPs-HN。结果表明，植物产生的 MRFV-VLPs 具有作为表面配体多价展示的新型平台的功能。
