Modified DNA aptamers against sweet agent aspartame.
We obtained a modified DNA aptamer against sweetener, aspartame, by in vitro selection method. The modified DNA was prepared from dATP, dGTP, dCTP and a modified dTTP bearing a terminal amino group at C-5 position in place of thymidine by PCR using a hyper thermophilic DNA polymerase, KOD Dash DNA polymerase. The synthetic 102-mer DNA with a 60-mer random region was used as an initial template for the PCR. The PCR-amplified modified DNA library was applied to an aspartame-agarose column, and then the bound modified DNA was eluted from the column for the affinity chromatography selection. Repeating the procedure, we selected the modified DNA aptamer against aspartame.
Saitoh
Hiroshi
H
Department of Applied Chemistry, Faculty of Engineering, Gunma University, Kiryu, Gunma 376-8515, Japan.
Nakamura
Akiko
A
Kuwahara
Masayasu
M
Ozaki
Hiroaki
H
Sawai
Hiroaki
H
D001218:Aspartame / Q000737:chemistry*; D001483:Base Sequence; D004247:DNA / Q000737:chemistry*; D017931:DNA Primers; D016133:Polymerase Chain Reaction; D013549:Sweetening Agents / Q000737:chemistry*
D017931:DNA Primers; D013549:Sweetening Agents; D004247:DNA; D001218:Aspartame