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PMID: 1121421 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

Detection by electron microscopy of photo-induced denaturation in lambda DNA.

Nucleic acids research ·Vol. 2 ·No. 2 ·1975-02-00 ·Pages 143-8

Stafford RS, Allison DP, Rahn RO

Abstract

We have used an electron microscope to study localized denatured regions in ultraviolet-irradiated DNA. DNA from bacteriophage lambda was UV-irradiated and then prepared for electron microscopy after fixing in buffered (pH 9.5) formaldehyde solutions at 25 degrees C. The denatured regions observed corresponded to those described by Inman and Schnös (1) who used alkaline denaturation to preferentially destroy thymine-adenine base pairing. In UV-irradiated DNA, pairs of neighboring thymine residues are converted into photodimers; hence, loss of hydrogen bonding most likely occurs in thymine-rich regions and denaturation results. Conceivably, photo-induced denaturation may under some circumstances represent a more convenient method than alkaline denaturation for mapping thymine-rich regions in DNA.

MeSH Terms
Coliphages DNA Viruses DNA, Viral/radiation effects Microscopy, Electron Nucleic Acid Conformation Nucleic Acid Denaturation Silver Thymine/analysis Ultraviolet Rays
Chemicals
DNA, Viral Silver Thymine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Stafford R S
Allison D P
Rahn R O
References (8)
8 references, click to expand
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    Photochem Photobiol. 1974 Jan;19(1):75-8 PMID: 4590519
  4. Bacteriophage of Haemophilus influenzae. I. Differences between infection by whole phage, extracted phage DNA and prophage DNA extracted from lysogenic cells.
    J Mol Biol. 1972 Feb 14;63(3):335-48 PMID: 4536897
  5. Ultraviolet irradiation of nucleic acids complexed with heavy atoms. II. Phosphorescence and photodimerization of DNA complexed with Ag.
    Photochem Photobiol. 1973 Jul;18(1):29-38 PMID: 4744137
  6. Electron-microscopic visualization of deletion mutations.
    Proc Natl Acad Sci U S A. 1968 May;60(1):243-50 PMID: 5241526
  7. Effect of a single thymine photodimer on the oligodeoxythymidylate-polydeoxyadenylate interaction.
    J Am Chem Soc. 1971 Sep;93(19):4940-2 PMID: 5118217
  8. Mapping of deletions and substitutions in heteroduplex DNA molecules of bacteriophage lambda by electron microscopy.
    Science. 1969 Mar 21;163(3873):1343-8 PMID: 5765116
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/2.2.143
Published
1975-02-00
Pages
143-8
Language
English
Region
England
NLM ID
0411011
PMCID
PMC342821
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/2.2.143 PMC Full Text View journal details
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