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In vitro selection and characterization of single stranded DNA aptamers for luteolin: A possible recognition tool

Sabah, Jinan Tuma and Mohamed Zulkifli, Razauden and Shahir, Shafinaz and Ahmed, Farediah and Abdul Kadir, Mohammed Rafiq and Zakaria, Zarita (2018) In vitro selection and characterization of single stranded DNA aptamers for luteolin: A possible recognition tool. Analytical Biochemistry, 549 . pp. 72-79. ISSN 0003-2697

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Official URL: http://dx.doi.org/10.1016/j.ab.2018.03.004

Abstract

Distinctive bioactivities possessed by luteolin (3′ 4′ 5, 7-tetrahydroxy-flavone) are advantageous for sundry practical applications. This paper reports the in vitro selection and characterization of single stranded-DNA (ssDNA) aptamers, specific for luteolin (LUT). 76-mer library containing 1015 randomized ssDNA were screened via systematic evolution of ligands by exponential enrichment (SELEX). The recovered ssDNA pool from the 8th round was amplified with unlabeled primers and cloned into PSTBlue-1 vector prior to sequencing. 22 of LUT-binding aptamer variants were further classified into one of the seven groups based on their N40 random sequence regions, wherein one representative from each group was characterized. The dissociation constant of aptamers designated as LUT#28, LUT#20 and LUT#3 was discerned to be 107, 214 and 109 nM, respectively with high binding affinity towards LUT. Prediction analysis of the secondary structure suggested discrete features with typical loop and stem motifs. Furthermore, LUT#3 displayed higher specificity with insignificant binding toward kaempferol and quercetin despite its structural and functional similarity compared to LUT#28 and LUT#20. Further LUT#3 can detect free luteolin within 0.2–1 mM in solution. It was suggested that LUT#3 aptamer were the most suitable for LUT recognition tool at laboratory scale based on the condition tested.

Item Type:Article
Uncontrolled Keywords:Aptamer, Luteolin
Subjects:Q Science > Q Science (General)
Divisions:Biosciences and Medical Engineering
ID Code:85913
Deposited By: Widya Wahid
Deposited On:30 Jul 2020 15:38
Last Modified:30 Jul 2020 15:38

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