Premium
Quantitative measurements on the duplex stability of 2,6‐diaminopurine and 5‐chloro‐uracil nucleotides using enzymatically synthesized oligomers
Author(s) -
Jörg D. Hoheisel,
Hans Lehrach
Publication year - 1990
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(90)81340-t
Subject(s) - oligomer , uracil , oligonucleotide , chemistry , nucleotide , dissociation (chemistry) , duplex (building) , stereochemistry , dissociation constant , base pair , dna , biochemistry , organic chemistry , gene , receptor
2,6‐Diaminopurine and 5‐chloro‐uracil 2'‐deoxynucleoside 5'‐triphosphates were synthesized from their 2'‐deoxynucleosides. Using a method of creating oligonucleotides by enzymatic primer extension, dodecanucleotides representing an Xba I/ Sal I site and the complementary Sal I/ Xba I site were generated containing these base modifications. Their duplex stability was quantitatively compared by thin‐layer chromatography to oligomers containing 2'‐deoxyadenosine and 2'‐deoxythymidine. The two unmodified oligomers already showed significant differences in dissociation temperature and binding equilibrium. Substitution with 5‐chloro‐2'‐deoxyuridine did not affect the dissociation temperature of either oligomer, the 2,6‐diaminopurine, however, led to an increase of 1.8°C or 1.5°C per modified base, respectively. While in the Xba I/ Sal I oligomer both base modifications changed the binding equilibrium, the 2,6‐diaminopurine by a factor of 1.32, the 5‐chloro‐uracil by 0.65, no such effect was found with the complementary oligomer.