基于寨卡病毒复制子的抗病毒药物筛选系统  

An Antiviral Drug Screening Platform Based on Zika Replicon

作　　者：刘娜 胡媛媛 陈凯丽 赵莹 易文富 朱春玉[1] 郑方亮 LIU Na;HU Yuan-yuan;CHEN Kai-li;ZHAO Ying;YI Wen-fu;ZHU Chun-yu;ZHENG Fang-liang(College of Life Sciences,Liaoning University,Shenyang 110036,Liaoning,China)

机构地区：[1]辽宁大学生命科学学院,中国辽宁沈阳110036

出　　处：《生命科学研究》2020年第2期95-101,117,共8页Life Science Research

基　　金：国家自然科学基金资助项目(31600128,31500131);辽宁省自然科学基金资助项目(20180551069)。

摘　　要：寨卡病毒(Zika virus,ZIKV)是引起成人格林巴利综合征(Guillain-Barre syndrome,GBS)以及胎儿和新生儿小头畸形疾病的重要病原体。有效的抗病毒药物筛选系统是防治ZIKV感染的有效工具。本研究构建了以海肾荧光素酶为检测靶标的寨卡病毒复制子Rluc-ZIKV-Rep,并通过突变NS5蛋白,构建了非复制型寨卡病毒复制子Rluc-ZIKV-Rep NS5△GDD。为了检测复制子的功能,将两种复制子转化至Vero细胞,结果显示随着时间延长,Rluc-ZIKV-Rep荧光素酶活性逐渐增加,寨卡病毒mRNA拷贝数逐渐增多,证明以海肾荧光素酶为检测靶标的寨卡病毒复制子构建成功。为了检测该系统是否能够用于抗病毒药物筛选,用马尼地平和西尼地平两种抗寨卡病毒的药物进行检测,结果显示Rluc-ZIKV-Rep的复制能力随着药物浓度的增加而逐渐降低,呈现一种剂量依赖的方式,说明该系统可以用于抗病毒药物的筛选。综上所述,本研究构建了一种可以用于抗病毒药物筛选的寨卡病毒复制子,为筛选有效的抗病毒药物提供了有力工具。Zika virus(ZIKV) is an important pathogen that causes Guillain-Barre syndrome(GBS) of adults and microcephaly of babies. The antiviral drug screening system is an effective tool for preventing and treat-ing ZIKV infection. Herein, Rluc-ZIKV-Rep, a ZIKV replicon with Renilla luciferase as a detection target,was constructed, and a non-replicating ZIKV replicon Rluc-ZIKV-Rep NS5△GDD was constructed by mutating NS5 protein. In order to test their functions, the two replicons were transformed into Vero cells, and it was found that the Rluc-ZIKV-Rep luciferase activity and the ZIKV mRNA copy numbers gradually in-creased over time, proving that ZIKV replicon with Renilla luciferase as an indicator was successfully con-structed. In order to test whether the system can be used for antiviral drug screening, two anti-ZIKV drugs,manidipine and cilnidipine were used in the test, and it was found that the replication ability of Rluc-ZIKV-Rep gradually reduced with the increasing drug concentration in a dose-dependent manner, indicat-ing that the system can be used for antiviral drug screening. In summary, a ZIKV replicon that can be used for antiviral drug screening was constructed successfully, and it would be a powerful tool for screening effective antiviral drugs.

关 键 词：寨卡病毒 复制子 抗病毒药物筛选 海肾荧光素酶 病毒复制 

分 类 号：Q782[生物学—分子生物学]