Key Laboratory of Marine Bio-resources Sustainable Utilization, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, PR China.
Virus Res. 2011 Sep;160(1-2):221-9. doi: 10.1016/j.virusres.2011.06.018. Epub 2011 Jul 2.
Green fluorescent protein-tagged recombinant virus has been successfully applied to observing the infective dynamics and evaluating viral replication. Here, we identified soft-shelled turtle iridovirus (STIV) ORF55 as an envelope protein (VP55), and developed a recombinant STIV expressing an enhanced green fluorescent protein (EGFP) fused to VP55 (EGFP-STIV). Recombinant EGFP-STIV shared similar single-step growth curves and ultrastructural morphology with wild type STIV (wt-STIV). The green fluorescence distribution during EGFP-STIV infection was consistent with the intracellular distribution of VP55 which was mostly co-localized with virus assembly sites. Furthermore, EGFP-STIV could be used to evaluate viral replication conveniently under drug treatment, and the result showed that STIV replication was significantly inhibited after the addition of antioxidant pyrrolidine dithiocarbamate (PDTC). Thus, the EGFP-tagged recombinant iridovirus will not only be useful for further investigations on the viral replicative dynamics, but also provide an alternative simple strategy to screen for antiviral substances.
绿色荧光蛋白标记的重组病毒已成功应用于观察感染动力学和评估病毒复制。在这里，我们鉴定了中华鳖虹彩病毒（STIV）ORF55 作为一种包膜蛋白（VP55），并开发了一种表达与 VP55 融合的增强型绿色荧光蛋白（EGFP）的重组 STIV（EGFP-STIV）。重组 EGFP-STIV 与野生型 STIV（wt-STIV）具有相似的单步生长曲线和超微结构形态。在 EGFP-STIV 感染过程中绿色荧光的分布与 VP55 的细胞内分布一致，VP55 主要与病毒装配部位共定位。此外，EGFP-STIV 可在药物处理下方便地用于评估病毒复制，结果表明，抗氧化剂吡咯烷二硫代氨基甲酸盐（PDTC）的加入显著抑制了 STIV 的复制。因此，该 EGFP 标记的重组虹彩病毒不仅将有助于进一步研究病毒复制动力学，还为筛选抗病毒物质提供了一种替代的简单策略。
