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PMID: 345243 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Electron microscopic visualization of tRNA genes with ferritin-avidin: biotin labels.

Nucleic acids research ·Vol. 5 ·No. 2 ·1978-02-00 ·Pages 363-84

Broker TR, Angerer LM, Yen PH, Hershey ND, Davidson N

Abstract

A method is described for indirect electron microscopic visualization and mapping of tRNA and other short transcripts hybridized to DNA. This method depends upon the attachment of the electron-dense protein ferritin to the RNA, the binding being mediated by the remarkably strong association of the egg white protein avidin with biotin. Biotin is covalently attached to the 3' end of tRNA using an NH2(CH2)5NH2 bridge. The tRNA-biotin adduct is hybridized to complementary DNA sequences present in a single stranded non-homology loop of a DNA:DNA heteroduplex. Avidin, covalently crosslinked to ferritin, is mixed with the heteroduplex and becomes bound to the hybridized tRNA-biotin. Observation of the DNA:RNA-biotin:avidin-ferritin complex by electron microscopy specifically and accurately reveals the position of the tRNA gene, with a frequency of labeling of approximately 50%.

MeSH Terms
Avidin Biotin Chemical Phenomena Chemistry Chromosome Mapping Coliphages DNA, Bacterial/genetics Escherichia coli/genetics Ferritins Genes Microscopy, Electron/methods RNA, Bacterial/genetics RNA, Transfer/genetics
Chemicals
DNA, Bacterial RNA, Bacterial Avidin Biotin Ferritins RNA, Transfer
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Broker T R
Angerer L M
Yen P H
Hershey N D
Davidson N
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34 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/5.2.363
Published
1978-02-00
Pages
363-84
Language
English
Region
England
NLM ID
0411011
PMCID
PMC341989
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/5.2.363 PMC Full Text View journal details
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