Molecular cloning of hepatitis delta virus RNA from an infected woodchuck liver: sequence, structure, and applications.
cDNA prepared from the single-stranded circular RNA genome of hepatitis delta virus was cloned in lambda gt11 by using RNA from the liver of an infected woodchuck. From the sequence of overlapping clones, we assembled the full sequence of 1,679 nucleotides. The sequence indicated an exceptional ability for intramolecular base pairing, yielding a rod structure with at least 70% of the bases paired and a predicted free energy of -805 kcal (-3,368 kJ)/mol. Three of the lambda clones contained sequences that were not only expressed as fusion proteins with beta-galactosidase but were recognized by human hepatitis delta virus-specific antibody. These clones were sequenced so as to establish the reading frame of the delta antigen on the antigenomic strand. The fusion protein produced by one clone was purified by immunoaffinity chromatography and then was used to raise rabbit antibodies specific for the delta antigen.
Kuo
M Y
MY
Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111.
Goldberg
J
J
Coates
L
L
Mason
W
W
Gerin
J
J
Taylor
J
J
D000818:Animals; D000956:Antigens, Viral / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005796:Genes; D005814:Genes, Viral; D003698:Hepatitis Delta Virus / Q000235:genetics* / Q000276:immunology; D006860:Hydrogen Bonding; D008099:Liver / Q000382:microbiology; D008392:Marmota; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000276:immunology; D014764:Viral Proteins / Q000235:genetics
D000956:Antigens, Viral; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D004247:DNA