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	Generation and characterization of west nile pseudo-infectious reporter virus for antiviral screening
	Zhang, Hong-Lei1,2; Ye, Han-Qing1; Deng, Cheng-Lin1; Liu, Si-Qing1; Shi, Pei-Yong3,4; Qin, Cheng-Feng5; Yuan, Zhi-Ming1; Zhang, Bo1
刊名	Antiviral research
	2017-05-01
卷号	141页码:38-47
关键词	West nile virus Pseudo-infectious virus Reporter virus Bsl-3 Antiviral screening
ISSN号	0166-3542
DOI	10.1016/j.antiviral.2017.02.006
通讯作者	Zhang, bo(zhangbo@wh.iov.cn)
英文摘要	West nile virus (wnv), a mosquito-borne flavivirus, is an important neurotropic human pathogen. as a biosafety level-3 (bsl-3) agent, wnv is strictly to bsl-3 laboratories for experimentations, thus greatly hindering the development of vaccine and antiviral drug. here, we developed a novel pseudo-infectious wnv reporter virus expressing the gaussia luciferase (gluc). a stable 293tnsi cell line expressing ns1 was selected for trans-supplying ns1 protein to support the replication of wnv-ans1 virus and wnvans1-gluc reporter virus with large-fragment deletion of ns1. wnv-ans1 virus and wnv-gluc-ans1 reporter virus were confined to complete their replication cycle in this 293tns1 cell line, displaying nearly identical growth kinetics to wt wnv although the viral titers were lower than those of wt wnv. the reporter gene was stably maintained in virus genome at least within three rounds of passage in 293tnsi cell line. using a known flaviviruses inhibitor, nitd008, we demonstrated that the pseudo infectious wnv-gluc-ans1 could be used for antiviral screening. furthermore, a high-throughput screening (hts) assay in a 96-well format was optimized and validated using several known wnv inhibitors, indicating that the optimized hts assay was suitable for high-throughput screening wnv inhibitors. our work provides a stable and safe tool to handle wnv outside of a bsl-3 facility and facilitates high throughput screening for anti-wnv drugs. (c) 2017 elsevier b.v. all rights reserved.
WOS关键词	BORNE ENCEPHALITIS-VIRUS ; NONSTRUCTURAL PROTEINS NS1 ; DENGUE VIRUS ; RNA REPLICATION ; UNITED-STATES ; GAUSSIA LUCIFERASE ; VIRAL-RNA ; IN-VITRO ; PARTICLES ; NEUTRALIZATION
WOS研究方向	Pharmacology & Pharmacy ; Virology
WOS类目	Pharmacology & Pharmacy ; Virology
语种	英语
出版者	ELSEVIER SCIENCE BV
WOS记录号	WOS:000399506200005
内容类型	期刊论文
URI标识	http://www.corc.org.cn/handle/1471x/2373441
专题	武汉病毒研究所
通讯作者	Zhang, Bo
作者单位	1.Chinese Acad Sci, Wuhan Inst Virol, Key Lab Special Pathogens & Biosafety, Wuhan 430071, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 3.Univ Texas Med Branch, Sealy Ctr Struct Biol Mol Biophys, Dept Biochem & Mol Biol, Galveston, TX 77555 USA 4.Univ Texas Med Branch, Sealy Ctr Struct Biol Mol Biophys, Dept Pharmacol & Toxicol, Galveston, TX 77555 USA 5.Beijing Inst Microbiol & Epidemiol, State Key Lab Pathogen & Biosecur, Beijing 100071, Peoples R China
推荐引用方式 GB/T 7714	Zhang, Hong-Lei,Ye, Han-Qing,Deng, Cheng-Lin,et al. Generation and characterization of west nile pseudo-infectious reporter virus for antiviral screening[J]. Antiviral research,2017,141:38-47.
APA	Zhang, Hong-Lei.,Ye, Han-Qing.,Deng, Cheng-Lin.,Liu, Si-Qing.,Shi, Pei-Yong.,...&Zhang, Bo.(2017).Generation and characterization of west nile pseudo-infectious reporter virus for antiviral screening.Antiviral research,141,38-47.
MLA	Zhang, Hong-Lei,et al."Generation and characterization of west nile pseudo-infectious reporter virus for antiviral screening".Antiviral research 141(2017):38-47.

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