Invention Grant
US09052321B2 Flavivirus-based system for production of hepatitis C virus (HCV)
有权
用于生产丙型肝炎病毒(HCV)的基于黄病毒病毒的系统
- Patent Title: Flavivirus-based system for production of hepatitis C virus (HCV)
- Patent Title (中): 用于生产丙型肝炎病毒(HCV)的基于黄病毒病毒的系统
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Application No.: US13122154Application Date: 2009-09-28
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Publication No.: US09052321B2Publication Date: 2015-06-09
- Inventor: Bertrand Saunier , Miriam Triyatni , Edward A. Berger
- Applicant: Bertrand Saunier , Miriam Triyatni , Edward A. Berger
- Applicant Address: US DC Washington FR Paris
- Assignee: The United States of America, as represented by the Secretary, Department of Health and Human Services,INSERM (Institut National de la Sante et de la Recherche Medicale)
- Current Assignee: The United States of America, as represented by the Secretary, Department of Health and Human Services,INSERM (Institut National de la Sante et de la Recherche Medicale)
- Current Assignee Address: US DC Washington FR Paris
- International Application: PCT/US2009/058598 WO 20090928
- International Announcement: WO2010/039649 WO 20100408
- Main IPC: C12N5/10
- IPC: C12N5/10 ; C12N15/85 ; G01N33/68 ; C12Q1/68 ; G01N33/576 ; C07K14/005

Abstract:
Provided herein is a mammalian cell transformed to contain a plasmid encoding a T7 or SP6 promoter operably linked to one or more HCV genes, a subgenomic replicon from a flavivirus and a cytoplasmic T7 and SP6 RNA amplification system. Also provided herein are isolated replication-competent HCV particles produced by the method comprising the steps of providing a transformed mammalian cell according to the first embodiment, culturing the cell, and recovering the replication-competent HCV particles from the cell culture. Provided herein are isolated HCV structural proteins produced by the method comprising the steps of providing a transformed mammalian cell according to the first embodiment, culturing the cell, and recovering the HCV structural proteins from the cell culture. Further provided herein is a system for assaying HCV entry into a cell comprising a first plasmid encoding a T7 or SP6 promoter operably linked to an HCV polynucleotide comprising at least the 5′-UTR to NS2 operably linked to an EMCV IRES in frame with an SP6 or T7 polymerase gene, respectively, a first host cell line expressing a replicon from a flavivirus and comprising a cytoplasmic T7 and SP6 RNA amplification system, a second plasmid encoding a reporter gene operably linked to both T7 and SP6 promoters in tandem, and a second host cell line comprising a cytoplasmic T7 polymerase or SP6 polymerase RNA amplification system.
Public/Granted literature
- US20120100574A1 FLAVIVIRUS-BASED SYSTEM FOR PRODUCTION OF HEPATITIS C VIRUS (HCV) Public/Granted day:2012-04-26
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