期刊
ANTIVIRAL RESEARCH
卷 150, 期 -, 页码 148-154出版社
ELSEVIER
DOI: 10.1016/j.antiviral.2017.12.017
关键词
Flavivirus; Zika virus; Luciferase; Antiviral screening; Replicon cell line
资金
- National Key Research and Development Program of China [2016YFD0500400, 2016YFC1200400]
- National Natural Science Foundation of China [81572003]
- Shenzhen science and technology program [CXZZ20150504163004339]
Zika virus (ZIKV) is an important emerging human pathogen associated with microcephaly, Guillain-Barre syndrome and meningoencephalitis. Developing rapid and reliable HTS assay is important for ZIKV drug discovery. Here, we constructed a dicistronic ZIKV replicon (ZIKV-Pac-Rluc-Rep) that contained the Renilla luciferase (Rluc) reporter gene separated from the puromycin N-acetyl-transferase (Pac) selectable marker by a short peptide cleavage site. A clonal replicon cell line stably expressing high level of ZIKV replicon was established by selection with puromycin. By optimizing cell number, compound concentration and incubation time, a robust replicon cell-based HTS assay was developed with a calculated Z' value of > 0.5. The fully optimized assay was further validated using several known flavivirus replication inhibitors. Altogether, the replicon cell-based HTS assay developed in this study will facilitate the discovery of antiviral compounds against ZIKV.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据