CaltechAUTHORS
Published October 1, 1983 | Published
Journal Article Open

Molecular cloning and characterization of hepatitis A virus cDNA

Ticehurst, John R.
Racaniello, Vincent R.
Baroudy, Bahige M.
Baltimore, David ORCID icon
Purcell, Robert H.
Feinstone, Stephen M.
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Abstract

Double-stranded cDNA was synthesized from hepatitis A virus (HAV) RNA and inserted into the Pst I site of pBR322. Restriction endonuclease digestion and cross-hybridization of fragments yielded a map of overlapping cloned cDNAs that included at least 99% of the viral genome. Molecular clones containing HAV cDNA were identified by hybridizing cloned cDNA to electrophoretically resolved RNA from uninfected and HAV-infected tissue culture cells. Cloned cDNA probes specifically hybridized to RNA from infected cells, and the predominant species identified had the characteristic genomic length of picornaviral RNA (~7,500 nucleotides). A partial sequence from the 3' end of the genome revealed 414 bases in an open reading frame followed by two closely spaced stop codons, a 60-base noncoding region, and a tract of poly(A).

Additional Information

© 1983 by the National Academy of Sciences. Communicated by Robert M. Chanock, June 30, 1983. The support and contributions of Charles Buckler, Robert Chanock, Roger Deeley, Dave Hoggan, Bill Hoyer, Ching-Juh Lai, Tom Miele, Kathy Mihalik, Terry Popkin, Arnold Rabson, Johnna Sears, Margaret Stewart, and especially Taylor Chestnut are greatly appreciated. We also thank Linda Jordan for editorial assistance. This work was supported by Grant AI-08388 from the National Institute of Allergy and Infectious Diseases and Grant CA-14051 from the National Cancer Institute (core grant to S. Luria). V.R.R. was supported by a postdoctoral fellowship from the National Institute of Allergy and Infectious Diseases. D.B. is an American Cancer Society Research Professor. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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