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Enzymatic sequence‐specific spin labeling of a DNA fragment containing the recognition sequence of Eco RI endonuclease
Author(s) -
Bobst A.M.,
Pauly G.T.,
Keyes R.S.,
Bobst E.V.
Publication year - 1988
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(88)80578-7
Subject(s) - oligonucleotide , nucleic acid , recognition sequence , dna , chemistry , polynucleotide , restriction enzyme , stereochemistry , site directed spin labeling , sequence (biology) , base pair , endonuclease , nuclear magnetic resonance spectroscopy , binding site , duplex (building) , spin label , covalent bond , crystallography , biochemistry , organic chemistry , membrane
Deoxyuridine analogs spin labeled in position 5 have been enzymatically incorporated sequence specifically into an oligodeoxyribonucleotide to form a spin‐labeled 26‐mer. The 26‐mer contains the Eco RI‐binding site and two labels which are located symmetrically close to the binding site. The labels are separated from one another far beyond the Heisenberg spin‐exchange distance. The local base motion as determined by ESR spectroscopy is of the order of 4 ns in the oligonucleotide duplex. This is the same value as reported earlier for local T motions in polynucleotide duplexes, thereby providing direct experimental evidence that the ESR line shape of spin levels covalently attached to nucleic acids depends primarily on the local dynamics of the nucleic acid building blocks.