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PMID: 6300836 Published · ppublish English Journal Article

Molecular cloning of double-stranded RNA virus genomes.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 80 ·No. 2 ·1983-01-00 ·Pages 373-7

Imai M, Richardson MA, Ikegami N, Shatkin AJ, Furuichi Y

Abstract

Genome double-stranded RNAs isolated from purified human reovirus (serotype 3) and rotavirus (Wa strain) were modified at the 3' termini by addition of oligo(C) approximately 15 with T4 RNA ligase. These RNAs were transcribed into cDNA by oligo(dG)-primed reverse transcriptase and cloned after insertion into pBR322 at the Pst I site. Hybridization of plasmid-transformed Escherichia coli RR1 colonies with 32P-labeled viral genome RNAs demonstrated the presence of DNA clones representative of each of the 10 reovirus RNAs and 10 of the 11 constituent segments of the rotavirus genome. Analyses of the size and terminal nucleotide sequences of insert DNAs indicated that some clones contained a full-length copy of the virus genome segment. The complete nucleotide sequence of rotavirus genome segment 11 double-stranded RNA was obtained by using this procedure. It provides a general method for cloning double-stranded RNAs and also nonpolyadenylylated single-stranded RNAs.

MeSH Terms
Amino Acid Sequence Base Sequence Cloning, Molecular DNA/metabolism Genes, Viral Humans Mammalian orthoreovirus 3/genetics Nucleic Acid Hybridization RNA, Double-Stranded/genetics RNA, Viral/genetics Reoviridae/genetics Rotavirus/genetics Transcription, Genetic Viral Proteins/genetics
Chemicals
RNA, Double-Stranded RNA, Viral Viral Proteins DNA
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Imai M
Richardson M A
Ikegami N
Shatkin A J
Furuichi Y
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20 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.80.2.373
Published
1983-01-00
Pages
373-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC393379
Subset
IM
Databases
GENBANK
J02440
External Links
PubMed source DOI: 10.1073/pnas.80.2.373 PMC Full Text View journal details
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