Publications [#262891] of Tuan Vo-Dinh

Papers Published

1. Giannetti, A; Citti, L; Domenici, C; Tedeschi, L; Baldini, F; Wabuyele, MB; Vo-Dinh, T, FRET-based protein-DNA binding assay for detection of active NF-κB, Sensors and Actuators, B: Chemical, vol. 113 no. 2 (February, 2006), pp. 649-654, Elsevier BV, ISSN 0925-4005 [014], [doi]
   (last updated on 2024/04/18)

   Abstract:
   A novel method to detect the active form of NF-κB, a transcription factor regulating a battery of inflammatory genes and playing a fundamental role in the development of numerous pathological states, has been developed. In the present work, we used fluorescence resonance energy transfer (FRET) to study DNA-protein binding interaction taking place between double-strand (ds) DNA immobilized in a glass capillary wall and p50 proteins. For this purpose, we developed a regenerable FRET-based system comprising of a single-strand (ss) DNA with auto-complementary sequence that is end-labeled with Cy5 dye and is highly specific for p50 proteins. The proteins were labeled with a Black Hole Quencher (BHQ-3) to be used as FRET pair. The interaction of p50/p50 homodimer active form with its DNA binding site was demonstrated by both electrophoretic mobility shift assays and FRET studies. These preliminary results demonstrated the feasibility of the FRET-based DNA technique to detect the active form of NF-κB protein with 90% detection efficiency. In addition, we show that the system is stable and highly regenerable. © 2005 Elsevier B.V. All rights reserved.

   Keywords:
   Proteins;Energy transfer;Genes;DNA sequences;Fluorescence;