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PMID: 4943795 Published · ppublish English Journal Article

Purification and in vitro transcription of a transfer RNA gene.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 68 ·No. 9 ·1971-09-00 ·Pages 2268-72

Daniel V, Beckmann JS, Sarid S, Grimberg JI, Herzberg M, Littauer UZ

Abstract

A gene specifying tyrosine transfer RNA has been purified and transcribed in vitro. The purification procedure made use of two specialized transducing phages carrying the tRNA(Tyr) gene of Escherichia coli inserted into their DNA in opposite orientations. The separated heavy strands of the two phages were annealed and the single-stranded tails of the resulting hybrid were removed by digestion with Neurospora endonuclease. The size of the purified double-stranded structures was determined by electron microscopy. These isolated duplexes served as template for the in vitro transcription of tRNA(Tyr)-like molecules.

MeSH Terms
Centrifugation, Density Gradient Coliphages DNA, Bacterial/isolation & purification Deoxyribonucleases Escherichia coli Genes Genetic Code Microscopy, Electron Molecular Weight Nucleic Acid Hybridization RNA, Transfer/biosynthesis Transduction, Genetic Tyrosine
Chemicals
DNA, Bacterial Tyrosine RNA, Transfer Deoxyribonucleases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Daniel V
Beckmann J S
Sarid S
Grimberg J I
Herzberg M
Littauer U Z
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22 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.68.9.2268
Published
1971-09-00
Pages
2268-72
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC389398
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.68.9.2268 PMC Full Text View journal details
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