Functional mapping of the genome of the B19 (human) parvovirus by in vitro translation after negative hybrid selection.
We have analyzed the coding capacity of B19 parvovirus transcripts by in vitro translation using the negative hybrid selection technique. Five different antisense oligonucleotides (18-mers) corresponding to different portions of the B19 genome were hybridized to RNA samples extracted from human erythroid bone marrow cells infected with B19 parvovirus in vitro, and RNase H was added to cleave specific B19 RNA molecules at selected sites. B19-specific translation products of these RNA samples were determined by immunoprecipitation. We localized the B19 nonstructural protein to the left-side transcript and the two capsid proteins to overlapping transcripts from the right side of the genome.
Ozawa
K
K
Cell Biology Section, National Heart, Lung, and Blood Institute, Bethesda, Maryland 20892.
Ayub
J
J
Young
N
N
D004900:Erythroblasts / Q000378:metabolism; D005814:Genes, Viral; D006801:Humans; D009693:Nucleic Acid Hybridization; D009838:Oligodeoxyribonucleotides / Q000378:metabolism; D010321:Parvoviridae / Q000235:genetics*; D014176:Protein Biosynthesis; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics
D009838:Oligodeoxyribonucleotides; D012367:RNA, Viral; D014764:Viral Proteins