[1]王美月,张振宇,王晓钧*.稳定表达人、马Tetherin蛋白的 MDCK细胞系的构建及其限制流感病毒活性的研究[J].中国预防兽医学报,2018,(07):626-629.[doi:0.3969/j.issn.1008-0589.201712023]
 WANG Mei-yue,ZHANG Zhen-yu,WANG Xiao-jun*.Evaluation of Tetherin’ s antiviral activity on its stably expressing MDCK cell lines[J].Chinese journal of preventive veterinary medicine,2018,(07):626-629.[doi:0.3969/j.issn.1008-0589.201712023]
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稳定表达人、马Tetherin蛋白的 MDCK细胞系的构建及其限制流感病毒活性的研究()
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2018年07
页码:
626-629
栏目:
免疫学
出版日期:
2018-08-15

文章信息/Info

Title:
Evaluation of Tetherin’ s antiviral activity on its stably expressing MDCK cell lines
文章编号:
1008-0589(2018)7-0626-04
作者:
 

王美月张振宇王晓钧*

 (中国农业科学院哈尔滨兽医研究所 兽医生物技术国家重点实验室/马传染病和慢病毒病研究创新团队,黑龙江 哈尔滨 150069)
Author(s):
WANG Mei-yue ZHANG Zhen-yu WANG Xiao-jun*
 (State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences,
Harbin 150069, China)
关键词:
人、马TetherinMDCK细胞系流感病毒
Keywords:
 human and equine Tetherin  MDCK cell lines  influenza virus
分类号:
S852.4
DOI:
0.3969/j.issn.1008-0589.201712023
文献标志码:
A
摘要:
为评估人、马Tetherin (huTetherin、eqTetherin)限制流感病毒的活性,本研究将huTetherin、eqTetherin克隆于慢病毒表达载体pLPCX中,分别构建重组质粒pLPCX-HA-huTetherin、pLPCX-HA-eqTetherin,并构建pLPCX-HA作为阴性对照。将上述构建的重组质粒,pVSV-G以及pcGP共转染293T细胞,获得假病毒后感染MDCK细胞,并利用嘌呤霉素筛选、纯化,建立稳定表达huTetherin和eqTetherin的MDCK细胞系。经western blot鉴定显示,该细胞系中huTetherin和eqTetherin可稳定表达。以反向遗传救获的马流感病毒(A/equine/Jilin/ 1/1989) (JL89)感染这两种细胞系,结果显示eqTetherin和huTetherin均对该流感病毒株具有显著限制作用。该研究为进一步评价和探索huTetherin和eqTetherin限制流感病毒的作用和机制奠定了基础。
 
Abstract:
To evaluate the antiviral activity of human and equine Tetherin ((huTetherin and eqTetherin), the huTetherin and eqTetherin genes were cloned to lentiviral expressing plasmid pLPCX, respectively. The recombinant plasmid, pVSV-G and pcGP were co-transfected into HEK293T cells so as to package pseudovirion. The pseudovirion was used to infect MDCK cells and cell lines stably expressing huTetherin and eqTetherin ((MDCK-huTetherin and MDCK-eqTetherin) were established using colony-purification in present of puromycin. The cell lines expressing huTetherin and eqTetherin were confirmed by western blot, indicating that huTetherin and eqTetherin were stably expressed in the MDCK-huTetherin and MDCK-eqTetherin cell lines. Moreover, the cell lines were used to determine restriction on influenza virus (A/equine/Jilin/1/1989), The results demonstrated that the influenza virus replication was significantly restrained by both the huTetherin and eqTetherin, when the virus was inoculated into the cell lines. These data would contribute to further study on restriction activity and mechanism of influenza virus infection.  
 

参考文献/References:

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[7]Yin Xin, Hu Zhe, Gu Qin-yong, et al. Equine tetherin blocks retrovirus release and its activity is antagonized by equine infectious anemia virus envelope protein [J]. J Virol, 2014, 88(2): 1259-1270.
[8]Mitchell R S, Katsura C, Skasko M A, et al. Vpu antagonizes BST-2-mediated restriction of HIV-1 release via β-TrCP and endo-lysosomal trafficking [J]. PLoS Pathog, 2009, 5(5): e1000450.
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                     (本文编辑:彭永刚)

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更新日期/Last Update: 2018-08-15