Reactivity of cytosine and thymine in single-base-pair mismatches with hydroxylamine and osmium tetroxide and its application to the study of mutations

Journal Article · Wed Jun 01 00:00:00 EDT 1988 · Proceedings of the National Academy of Sciences of the United States of America; (USA)

DOI:https://doi.org/10.1073/pnas.85.12.4397· OSTI ID:5413418

Cotton, R G.H.; Rodrigues, N R; Campbell, R D ^[1]

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1. Univ. of Oxford (England)

The chemical reactivity of thymine (T), when mismatched with the bases cytosine, guanine, and thymine, and of cytosine (C), when mismatched with thymine, adenine, and cytosine, has been examined. Heteroduplex DNAs containing such mismatched base pairs were first incubated with osmium tetroxide (for T and C mismatches) or hydroxylamine (for C mismatches) and then incubated with piperidine to cleave the DNA at the modified mismatched base. This cleavage was studied with an internally labeled strand containing the mismatched T or C, such that DNA cleavage and thus reactivity could be detected by gel electrophoresis. Cleavage at a total of 13 T and 21 C mismatches isolated (by at least three properly paired bases on both sides) single-base-pair mismatches was identified. All T or C mismatches studied were cleaved. By using end-labeled DNA probes containing T or C single-base-pair mismatches and conditions for limited cleavage, the authors were able to show that cleavage was at the base predicted by sequence analysis and that mismatches in a length of DNA could be readily detected by such an approach. This procedure may enable detection of all single-base-pair mismatches by use of sense and antisense probes and thus may be used by identify the mutated base and its position in a heteroduplex.

• OSTI ID: 5413418
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (USA), Vol. 85:12; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
DNA
STRAND BREAKS
DNA MISMATCH
CYTOCHEMISTRY
CYTOSINE
GENES
HYDROXYLAMINE
MUTATIONS
OSMIUM OXIDES
THYMINE
AMINES
AZINES
BIOCHEMISTRY
CHALCOGENIDES
CHEMISTRY
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANIC OXYGEN COMPOUNDS
OSMIUM COMPOUNDS
OXIDES
OXYGEN COMPOUNDS
PYRIMIDINES
REFRACTORY METAL COMPOUNDS
TRANSITION ELEMENT COMPOUNDS
URACILS
550201* - Biochemistry- Tracer Techniques