Title: | "Selection, Characterization, and Application of ssDNA Aptamer against Furaneol" |
Author(s): | Komarova N; Andrianova M; Glukhov S; Kuznetsov A; |
Address: | "Scientific-Manufacturing Complex Technological Centre, 1(-)7 Shokin Square, Zelenograd, 124498 Moscow, Russia. nat.v.kom@gmail.com. Scientific-Manufacturing Complex Technological Centre, 1(-)7 Shokin Square, Zelenograd, 124498 Moscow, Russia. smariika1987@gmail.com. Scientific-Manufacturing Complex Technological Centre, 1(-)7 Shokin Square, Zelenograd, 124498 Moscow, Russia. serglukhovmb@gmail.com. Scientific-Manufacturing Complex Technological Centre, 1(-)7 Shokin Square, Zelenograd, 124498 Moscow, Russia. kae@tcen.ru" |
DOI: | 10.3390/molecules23123159 |
ISSN/ISBN: | 1420-3049 (Electronic) 1420-3049 (Linking) |
Abstract: | "Furaneol is an aroma compound which occurs naturally in foods and is used as an artificial flavor. Detection of furaneol is required in food science and food processing industry. Capture- Systematic Evolution of Ligands by EXponential enrichment (SELEX) protocol was applied for the isolation of an aptamer binding to furaneol, a small volatile organic substance contributing to the flavor of various products. Thirteen cycles of selection were performed. The resulting DNA pool was cloned, using blunt-end cloning, and ninety-six plasmids were sequenced and analyzed. Eight oligonucleotides were selected as aptamer candidates and screened for the ability to bind to furaneol, using three different methods-magnetic-beads associated elution assay, SYBR Green I assay, and exonuclease protection assay. One of the candidates was further characterized as an aptamer. The apparent equilibrium constant was determined to be (1.1 +/- 0.4) microM, by the fluorescent method. The reported aptamer was applied for development of the ion-sensitive field-effect transistor (ISFET)-based biosensor, for the analysis of furaneol, in the concentration range of 0.1(-)10 microM" |
Keywords: | "Aptamers, Nucleotide/*chemistry/genetics Base Sequence Biosensing Techniques DNA, Single-Stranded/*chemistry Fluorescence Furans/*chemistry SELEX Aptamer Technique/*methods Capture-SELEX Isfet aptamer aptasensor furaneol;" |
Notes: | "MedlineKomarova, Natalia Andrianova, Mariia Glukhov, Sergey Kuznetsov, Alexander eng 16-19-10697/Russian Science Foundation/ Switzerland 2018/12/06 Molecules. 2018 Nov 30; 23(12):3159. doi: 10.3390/molecules23123159" |