Genetic analysis of the hepatitis C virus (HCV) genome from HCV-infected human T cells

K. Sugiyama, Nobuyuki Kato, T. Mizutani, Masanori Ikeda, T. Tanaka, K. Shimotohno

Research output: Contribution to journalArticle

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Abstract

We recently established a cell culture system for the replication of hepatitis C virus (HCV) by using a human T cell leukaemia virus type I-infected cell line, MT-2, and showed that the quasi-species of the hypervariable region 1 observed in the original inoculum became homogeneous in MT-2 cells 10 days after inoculation of HCV. In this study, we obtained HCV cDNA clones covering the whole viral genome by RT-nested PCR using RNA from HCV-infected cloned MT-2C cells, which support viral replication more efficiently, at 12 days after inoculation. A total of 41 distinct HCV cDNA clones covering almost the whole viral genome were also isolated from a cDNA library derived from the original inoculum. Molecular evolutional analyses comparing the sequences of the HCV clones obtained from both sources revealed that the HCV populations became homogeneous in more than half of the compared regions. This finding suggests that limited HCV populations are able to replicate in MT-2C cells. In addition, we isolated cDNA clones containing a 3' X-tail sequence, which was recently identified as a bona fide 3' terminus of the HCV genome, in the HCV-infected MT-2C cells and confirmed that the nucleotide sequence of the 3' X-tail was highly conserved, suggesting its implication in HCV replication. Finally, on the basis of the sequences of HCV cDNA clones obtained from HCV-infected MT-2C cells, we determined the entire nucleotide sequence of the HCV genome containing the 3' X-tail as a candidate for an infectious HCV molecular clone.

Original languageEnglish
Pages (from-to)329-336
Number of pages8
JournalJournal of General Virology
Volume78
Issue number2
Publication statusPublished - Feb 1997
Externally publishedYes

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Hepacivirus
Genome
T-Lymphocytes
Clone Cells
Complementary DNA
Viral Genome
Tail
Hepatovirus
Human T-lymphotropic virus 1
Virus Replication
Gene Library
Population
Sequence Analysis
Cell Culture Techniques

ASJC Scopus subject areas

  • Virology
  • Immunology

Cite this

Sugiyama, K., Kato, N., Mizutani, T., Ikeda, M., Tanaka, T., & Shimotohno, K. (1997). Genetic analysis of the hepatitis C virus (HCV) genome from HCV-infected human T cells. Journal of General Virology, 78(2), 329-336.

Genetic analysis of the hepatitis C virus (HCV) genome from HCV-infected human T cells. / Sugiyama, K.; Kato, Nobuyuki; Mizutani, T.; Ikeda, Masanori; Tanaka, T.; Shimotohno, K.

In: Journal of General Virology, Vol. 78, No. 2, 02.1997, p. 329-336.

Research output: Contribution to journalArticle

Sugiyama, K, Kato, N, Mizutani, T, Ikeda, M, Tanaka, T & Shimotohno, K 1997, 'Genetic analysis of the hepatitis C virus (HCV) genome from HCV-infected human T cells', Journal of General Virology, vol. 78, no. 2, pp. 329-336.
Sugiyama K, Kato N, Mizutani T, Ikeda M, Tanaka T, Shimotohno K. Genetic analysis of the hepatitis C virus (HCV) genome from HCV-infected human T cells. Journal of General Virology. 1997 Feb;78(2):329-336.
Sugiyama, K. ; Kato, Nobuyuki ; Mizutani, T. ; Ikeda, Masanori ; Tanaka, T. ; Shimotohno, K. / Genetic analysis of the hepatitis C virus (HCV) genome from HCV-infected human T cells. In: Journal of General Virology. 1997 ; Vol. 78, No. 2. pp. 329-336.
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