Enzymatic and chemical synthesis of isodeoxynucleic acid (INA) and its properties.
Isodeoxynucleic acid (INA) is one of the most attractive sugar modified nucleic acids for therapeutic use. We report here the enzymatic synthesis and properties of INA. All four isodeoxynucleoside triphosphates (INTPs) were synthesized for use in a single nucleotide insertion reaction. K(m) and V(max) values showed the almost same order as natural dNTPs. Primer extension reactions were also carried out to investigate the function of INTPs as substrates for several DNA polymerases. INTPs were incorporated into N + 2 position. One INTP and three dNTPs were able to generate a full-length product (27 mer). These results suggested that template-primer duplex containing one isodeoxynucleoside could be recognized by DNA polymerases. However, duplexs containing more than two isodeoxynucleosides could not be recognized by DNA polymerases because of conformational changes by isodeoxynucleosides.
Ogino
Takaaki
T
Graduate School of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, Japan.
Inoue
Naonori
N
Sato
Kousuke
K
Matsuda
Akira
A
D004247:DNA / Q000096:biosynthesis / Q000737:chemistry; D017931:DNA Primers; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D003854:Deoxyribonucleotides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism*; D007700:Kinetics; D052156:SELEX Aptamer Technique; D013698:Templates, Genetic
D017931:DNA Primers; D003854:Deoxyribonucleotides; D004247:DNA; D004259:DNA-Directed DNA Polymerase