Davidson Andrew D
School of Cellular and Molecular Medicine, University of Bristol, Bristol, BS8 1TD, UK,
Methods Mol Biol. 2014;1138:113-30. doi: 10.1007/978-1-4939-0348-1_8.
The development of dengue virus "reverse genetic" systems based on full-length cDNA clones corresponding to the viral RNA genome has been an important technological platform for advancing dengue virus research. Mutations can be introduced into the genome to study their effect on virus replication and pathogenesis while attenuated or chimeric viruses can be constructed that are potential vaccine candidates. The deletion of the virus structural genes has led to the production of noninfectious, but replication competent viral subgenomes (termed replicons) that have been used to study viral replication and are useful for the screening of antiviral compounds. This article describes the development of dengue virus reverse genetic systems and protocols to manipulate the viral genome, recover infectious virus, and produce replicon-containing cell lines.
基于与病毒RNA基因组相对应的全长cDNA克隆开发登革病毒“反向遗传”系统，已成为推进登革病毒研究的重要技术平台。可将突变引入基因组以研究其对病毒复制和发病机制的影响，同时可构建减毒或嵌合病毒，它们是潜在的疫苗候选物。病毒结构基因的缺失导致产生无感染性但具有复制能力的病毒亚基因组（称为复制子），这些复制子已被用于研究病毒复制，并且有助于筛选抗病毒化合物。本文描述了登革病毒反向遗传系统的开发以及操纵病毒基因组、回收感染性病毒和产生含复制子细胞系的方案。
