Summary
In order to clone hepatitis C (blood-borne non-A, non-B hepatitis) virus, lambda gt11cDNA 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, hepatitits 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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bradley DW, Maynard JE: Etiology and natural history of post-transfusion and enterically-transmitted non-A, non-B hepatitis. Seminars in Liver Disease 1986;6:56–66
Feinstone SM, Alter HJ, Dienes HP, et al: Non-A, non-B hepatitis in chimpanzees and marmosets. J Infec Dis 1981;144:588–589
Dienstag JL: Noft-A, non-B hepatitis. II. Experimental transmission, putative virus agents, markers, and prevention. Gastroenterology 1983;85:743–768
Berman M, Alter HJ, Ishak KG, et al: The chronic sequelae of non-A, non-B hepatitis. Ann Intern Med 1979;91:1–6
Alter HJ, Purcell RH, Feinstone SM, et al: Non-A, non-B hepatitis: a review and interim report of an ongoing prospective study. In: Vyas GN, Cohen SN, Schmidt T, eds. Viral hepatitis: etiology, epidemiology, pathogenesis and prevention. Franklin Institute Press, Philadelphia. 1978;359–369
Dienstag JL: Non-A, non-B hepatitis, i. Recognition, epidemiology, and clinical features. Gastroenterology 1983;85:439–462
Hakim AA: Isolation and functional property of mRNA coding for hepatitis A, B, and non-A, non-B viral particles from human sera. Naturwissenschaften 1986;73:45–47
Linke HK, Amann CW, Schlauder GG: Non-A, non-B hepatitis infection does not result in the production of abundant poly-A-containing messenger RNAs. In:Zuckerman AJ, ed. Viral hepatitis and liver disease. Alan R. Liss, INC,New York 1988;564–567
Chomczynski P, Sacchi N: Single-step method of RNA isolation by acid guanidium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987;162:156–159
Maniatis T, Fritsch EF, Sambrook J: DNase that is free of RNase. In: Maniatis T, Fritsch EF, Sambrook J, eds. Molecular cloning. Cold Spring Harbor, New York1982;451–452
Davis LG, Dibner MD, Battey JF: Agarose gel electrophoresis. In: Davis LG, Dibner MD, Battey JF, eds. Basic methods in quencing. ibid. 1986;233–274
Davis LG, Dibner MD, Battey JF: cDNA cloning into lambda gtlO and lambda gt11.ibid 1986;193–218
Davis LG, Dibner MD, Battey JF: M13 cloning and sequencing,ibid 1986;233–274
Hattori M, Sakaki Y: Dideoxy sequencing method using denatured plasmid templates. Anal Biochem 1986;152:232–238
Davis LG, Dibner MD, Battey JF: Preparation of DNA from eukaryotic cells. In: Davis LG, Dibner MD, Battey JF, eds. Basic methods in molecular biology. Elsevier Science Publishing Co., INC, Amsterdam. 1986;41–66
Sumiyoshi H, Mori C, Fuke I, et al: Complete nucleotide sequence of the Japanese encephalitis virus genome RNA. Virology 1987;161:497–510
Alter HJ: Acute and chronic consequences of transfusion associated hepatitis. In: The American Association for the Study of Liver Diseases, ed. Hepatology update-portal hypertension; viral hepatitis. AASLD, Chicago. 1986;220–228
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Arima, T., Nagashima, H., Murakami, S. et al. Cloning of a cDNA associated with acute and chronic hepatitis C infection generated from patients serum RNA. Gastroenterol Jpn 24, 540–544 (1989). https://doi.org/10.1007/BF02773882
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02773882