肠道病毒71型5'端非编码区双链寡核苷酸抗肠道病毒的研究.doc

文档介绍：
基于肠道病毒 71 型 5’端非编码区双链寡核
苷酸抗肠道病毒的研究#
董星辰,罗震,葛茂林,吴建国**
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(武汉大学生命科学学院病毒学国家重点实验室,武汉 430072)
摘要:【目的】研究双链寡核苷酸(dsON)在体外抗肠道病毒 71(EV71)的效应。【方法】
设计合成了针对 EV71 5‘UTR 茎环结构的双链寡核苷酸,克隆 EV71 5‘UTR 到荧光素酶报
告基因系统中检测寡核苷酸对 5‘UTR 的翻译的影响,采用实时荧光定量 PCR(Real-time
RT-PCR)的方法检测细胞内病毒的 RNA 水平,采用蛋白质印迹(Western Blotting)检测细
胞内病毒蛋白的表达水平。【结果】双链寡核苷酸可以显著抑制病毒蛋白的表达。【结论】
针对 5’UTR 茎环结构的双链寡核苷酸具有一定的抗 EV71 的作用,可能作为一种治疗 EV71
感染的新途径。
关键词:肠道病毒 71;双链寡核苷酸;抗病毒
Study on antiviral activity of double strands oligonucleotides
target at IRES in 5’UTR of Enterovirus 71
DONG xinchen, LUO zhen, GE maolin, WU jianguo
(State Key Laboratory of Virology, College of Life Sciences Wuhan University, WuHan 430072)
Abstract: Objective To find out the antiviral effects of double strands oligodnucleotides(dsON)
to EV71 in vitro. Method we designed and synthesized different dsONs targeted at stem-loops in
5’UTR of EV71. Their antiviral activity was evaluated by Luciferase Reporter Assay, Real-Time
PCR, and Western Blotting. Result dsONs could definitely inhibit EV71 both in viral RNA and
protein. Conclusion The synthesized dsONs has antiviral activity and might be a new way for
treating EV71 infection.
Keywords: Enterovirus 71; Oligodnucleotides; antiviral activity
0 引言
肠道病毒 71(Enterovirus 71)是引起年龄 5 岁以下儿童手足口病(hand-foot-and-mouth
Disease, HFMD)的主要病因之一,严重感染会导致无菌性脑膜炎、神经系统紊乱甚至死亡
[1]
核苷酸(oligonue