Rajendran K S, Nagy Peter D
Department of Plant Pathology, University of Kentucky, Lexington, KY 40546, USA.
Methods Mol Biol. 2008;451:267-77. doi: 10.1007/978-1-59745-102-4_19.
Replication of the viral RNA genome performed by the viral replicase is the central process during the viral infection cycle (Nagy and Pogany, see earlier chapter four). Most RNA viruses assign one or more proteins translated from their own genomes for assembling the viral replicase complex, which consists of the viral RNA, viral proteins, and several subverted host proteins embedded in cellular membranes. Understanding the various biochemical activities of the replication proteins can lead to target identification for human intervention to control viral infections or the damage to the host cells. The replicase proteins of tomato bushy stunt virus (TBSV) are selected as model system to study the dynamics of interactions between viral replicase proteins using surface plasmon resonance (SPR) analysis. The SPR assay provides real-time protein interaction data by measuring the change in refractive index at the surface of the sensor chip due to the change in mass resulting from the interaction between the immobilized protein and the protein that is being passed over the immobilized chip surface. SPR-based biosensor BIAcore X was used to carry out TBSV replicase protein interaction studies.
由病毒复制酶进行的病毒RNA基因组复制是病毒感染周期中的核心过程（纳吉和波加尼，见第四章前文）。大多数RNA病毒利用从其自身基因组翻译而来的一种或多种蛋白质来组装病毒复制酶复合体，该复合体由病毒RNA、病毒蛋白以及嵌入细胞膜中的几种被颠覆的宿主蛋白组成。了解复制蛋白的各种生化活性有助于确定人类干预以控制病毒感染或宿主细胞损伤的靶点。番茄丛矮病毒（TBSV）的复制酶蛋白被选作模型系统，以利用表面等离子体共振（SPR）分析来研究病毒复制酶蛋白之间相互作用的动态过程。SPR分析通过测量传感器芯片表面由于固定化蛋白与流经固定化芯片表面的蛋白之间相互作用导致的质量变化而引起的折射率变化，提供实时蛋白质相互作用数据。基于SPR的生物传感器BIAcore X用于开展TBSV复制酶蛋白相互作用研究。
