國家衛生研究院 NHRI:Item 3990099045/1149
English  |  正體中文  |  简体中文  |  Items with full text/Total items : 12145/12927 (94%)
Visitors : 852812      Online Users : 350
RC Version 6.0 © Powered By DSPACE, MIT. Enhanced by NTU Library IR team.
Scope Tips:
  • please add "double quotation mark" for query phrases to get precise results
  • please goto advance search for comprehansive author search
  • Adv. Search
    HomeLoginUploadHelpAboutAdminister Goto mobile version
    Please use this identifier to cite or link to this item: http://ir.nhri.org.tw/handle/3990099045/1149


    Title: A reporter-based assay for identifying hepatitis C virus inhibitors based on subgenornic replicon cells
    Authors: Lee, JC;Chang, CF;Chi, YH;Hwang, DR;Hsu, JTA
    Contributors: Division of Biotechnology and Pharmaceutical Research
    Abstract: Hepatitis C virus (HCV) encodes a polyprotein that needs to be processed proteolytically by cellular and viral proteases into mature functional proteins. One of the viral proteins, NS3/4A, has serine protease activity that is critical for virus maturation. The generation and characterization of an engineered HCV replicon cell line (Ava5) is described which constitutively expresses EG(Delta4AB)SEAP reporter protein and the cell line was designated as Ava5-EG(Delta4AB)SEAP.EG(Delta4AB)SEAP is a fusion protein in which Enhanced Green Fluorescent Protein (EGFP) was fused to SEcreted Alkaline Phosphatase (SEAP) through the NS3/4A protease decapeptide recognition sequence, Delta4AB, which spans the NS4A and NS4B junction region. The secretion of SEAP into culture medium has been shown to depend on the cleavage of Delta4AB by HCV NS3/4A protease. It is demonstrated that the amount of NS3/4A in Ava5-EG(Delta4AB)SEAP cells correlated well with the copy numbers of HCV subgenomic RNA. It is also shown that replication of HCV subgenomic RNA inside cells is reflected by the alkaline phosphatase (SEAP) levels in culture medium. SEAP activity in the culture medium of Ava5-EG(Delta4AB)SEAP was approximately 50-fold higher than the parental Ava5 cells. Ava5-EG(Delta4AB)SEAP was validated as a drug screening system since several known HCV inhibitors were shown to reduce SEAP activities in culture media of Ava5-EG(Delta4AB)SEAP cells. In conclusion, Ava5-EG(Delta4AB)SEAP cells can be used to monitor HCV sub-genomic replication and the assay can be readily adapted to high throughput screening format to identify prospective anti-HCV drugs. (C) 2003 Elsevier B.V. All rights reserved.
    Keywords: Biochemical Research Methods;Biotechnology & Applied Microbiology;Virology
    Date: 2004-03-01
    Relation: Journal of Virological Methods. 2004 Mar;116(1):27-33.
    Link to: http://dx.doi.org/10.1016/j.jviromet.2003.10.007
    JIF/Ranking 2023: http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=NHRI&SrcApp=NHRI_IR&KeyISSN=0166-0934&DestApp=IC2JCR
    Cited Times(WOS): https://www.webofscience.com/wos/woscc/full-record/WOS:000188380600004
    Cited Times(Scopus): http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=0346096567
    Appears in Collections:[John Tsu-An Hsu] Periodical Articles
    [Ya-Hui Chi] Periodical Articles

    Files in This Item:

    File Description SizeFormat
    000188380600004.pdf206KbAdobe PDF1073View/Open


    All items in NHRI are protected by copyright, with all rights reserved.

    Related Items in TAIR

    DSpace Software Copyright © 2002-2004  MIT &  Hewlett-Packard  /   Enhanced by   NTU Library IR team Copyright ©   - Feedback