Cloning of a cDNA associated with acute and chronic hepatitis C infection generated from patients serum RNA.
In order to clone hepatitis C (blood-borne non-A, non-B hepatitis) virus, lambda gt11-cDNA library was constructed from RNA extracted from 100 liters serum collected from 1,047 donors with elevated ALT levels and negative for hepatitis B virus-DNA. The library was immunoscreened on Y1090 cells with pooled serum obtained from patients with acute hepatitis C or chronic hepatitis C. By screening 29 clones specific for Japanese hepatitis C infection were isolated. The specificity of these clones for hepatitis C infection was determined by panels constructed in 3 laboratories. Of these, 12 clones were specific for American hepatitis C infection as well. The nucleotide sequence (201 bp) of one of them was determined to be unique compared to known human viruses including hepatitis A virus, hepatitis B virus and hepatitis D virus. Southern blot analysis showed the absence of the sequence of the human genome in the clone. The predicted amino acid sequence is rich in residues of lysine, arginine, glutamic acid and asparagine, while lacking leucine, cysteine and methionine.
Arima
T
T
First Department of Internal Medicine, Okayama University Medical School, Japan.
Nagashima
H
H
Murakami
S
S
Kaji
C
C
Fujita
J
J
Shimomura
H
H
Tsuji
T
T
D000208:Acute Disease; D000595:Amino Acid Sequence; D001483:Base Sequence; D002908:Chronic Disease; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics*; D006526:Hepatitis C / Q000097:blood / Q000235:genetics*; D006525:Hepatitis, Viral, Human / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000032:analysis / Q000097:blood*; D012689:Sequence Homology, Nucleic Acid
D004279:DNA, Viral; D012367:RNA, Viral; D004247:DNA