Sequence, expression and reconstitution of an HCV genome from a British isolate derived from a single blood donation.
Morphological analysis of hepatitis C virus and development of antiviral drugs to eradicate this agent have been seriously hampered by the low viraemias observed during natural infection and the unavailability of a cell culture system for virus propagation. Recently a low-grade hepatitis has been reported in chimpanzees after intrahepatic transfection of full-length synthetic HCV RNA and successful infections shown to be critically dependent on the integrity and genetic homogeneity of the reconstituted clone. In this study we describe and characterize a full HCV RNA sequence derived from a case of chronic sporadic hepatitis. The genotype was shown to be 1a with a low level of intraclonal sequence heterogeneity, and processing of both structural and nonstructural proteins has been documented. The assembly of the full genome has also been achieved. The low level of intraclonal variation observed may reflect infection with a single isolate and the fact that cloning was performed on virus obtained from a single blood donation makes this clone a good candidate for future in vivo and in vitro transfection studies.
Kumar
U
U
Department of Medicine, Imperial College School of Medicine, London, UK.
Tuthill
T
T
Thomas
H C
HC
Monjardino
J
J
D000818:Animals; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism; D001782:Blood Donors; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism; D016679:Genome, Viral; D016174:Hepacivirus / Q000235:genetics* / Q000302:isolation & purification; D019698:Hepatitis C, Chronic / Q000821:virology*; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D020815:Polyproteins / Q000378:metabolism; D014176:Protein Biosynthesis; D012367:RNA, Viral / Q000097:blood / Q000235:genetics; D018411:Spodoptera; D014158:Transcription, Genetic; D006113:United Kingdom; D014764:Viral Proteins / Q000378:metabolism
D004279:DNA, Viral; D020815:Polyproteins; D012367:RNA, Viral; D014764:Viral Proteins