國家衛生研究院 NHRI:Item 3990099045/1163

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Title:	Emergence of mutation clusters in the HCV genome during sequential viral passages in Sip-L expressing cells
Authors:	Hwang, DR;Lai, HY;Chang, ML;Hsu, JTA;Yeh, CT
Contributors:	Division of Biotechnology and Pharmaceutical Research
Abstract:	Sip-L, a member of the Cupin superfamily, is a hepatic factor capable of supporting hepatitis C virus (HCV) replication in an otherwise non-permissive cell line. HCV-positive serum was used to infect Huh-7 and 293 cells stably expressing Sip-L. Using the culture medium of the infected cells as an infection source, sequential viral passages were carried out in both cell lines. Efficient viral passage was observed in 293-Sip-L cells but not in Huh-7-Sip-L cells. The viral concentrations in the culture medium increased gradually from less than 10(2) copies/mL to 5.3 x 10(4) copies/mL after 25 sequential passages in 293-Sip-L cells. Sequence analysis of the viral genomes obtained from both the initial and final inocula revealed emergence of mutation. clusters in NS2, NS3, and NS5A coding regions. Immunofluorescence study revealed that only a small percentage of infected cells expressed a detectable level of viral protein. Caspase 3 activities in the infected cells increased progressively during the viral passages. In conclusion, perpetual propagation of HCV was achieved using Sip-L expressing cells, allowing for the development of mutation clusters in the genome. The mutant HCV can be used as an infection source to study the molecular mechanism of HCV replication.
Keywords:	Biochemical Research Methods;Biotechnology & Applied Microbiology;Virology
Date:	2005-11
Relation:	Journal of Virological Methods. 2005 Nov;129(2):170-177.
Link to:	http://dx.doi.org/10.1016/j.jviromet.2005.05.026
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:000232522400008
Cited Times(Scopus):	http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=25144484985
Appears in Collections:	[John Tsu-An Hsu] Periodical Articles

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