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Determination of DNA Binding Behavior of FoxA1 Constructs Using a Gold Nanoparticle-Based High Throughput Assay
Khin Moh Moh Aung1; Michelle Gek Liang Lim2; Shuzhen Hong2; Edwin Cheung2; Xiaodi Su1,3
2017-03-31
Source Publicationjournal of Molecular and Engineering Materials
ISSN2251-2373
Volume4Issue:3
Abstract

Forkhead box protein 1 (FoxA1) is a member of the forkhead family of winged-helix transcription factors. It plays crucial roles in the development and differentiation of multiple organs and in the regulation of estrogen-stimulated genes. In this study, in order to determine the regions of FoxA1 necessary for efficient Deoxyribonucleic Acid (DNA) binding, we cloned, expressed and purified a series of FoxA1 constructs that contain either the DNA Binding Domain (DBD), the Transcription Activation Domain (TAD), or both. We determined the DNA binding behavior of these constructs using traditional electrophoretic mobility shift assay (EMSA) and a recently developed gold nanoparticles (AuNPs)-based fast screening method. We conclude that just the DBD region alone is not sufficient for protein-DNA binding activity. Amino acids flanking the upstream of the DBD region are required for maximal DNA binding activity. Through this study, we have also further validated the AuNPs assay for its generality and expanded the existing protocol for comparing the DNA binding behavior of multiple proteins of different charge properties and molecular weights.

KeywordGold Nanoparticles Protein-dna Interactions Emsa Forkhead Box Protein Mutant Construct
DOIhttps://doi.org/10.1142/S2251237316400128
Indexed BySCI
Language英语
WOS Research AreaMaterials Science
WOS SubjectMaterials Science, Multidisciplinary
WOS IDWOS:000399159800005
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Document TypeJournal article
CollectionFaculty of Health Sciences
Corresponding AuthorEdwin Cheung; Xiaodi Su
Affiliation1.Institute of Materials Research and Engineering, Agency for Science Technology and Research (A*STAR) #08-03 Innovis, 2 Fusionopolis Way, Singapore 138634
2.Genome Institute of Singapore, Agency for Science Technology and Research (A*STAR) #02-01 Genome 60 Biopolis Street, Singapore 138672
3.Department of Chemistry, National University of Singapore Block S8, Level 3, 3 Science Drive, Singapore 117543
Recommended Citation
GB/T 7714
Khin Moh Moh Aung,Michelle Gek Liang Lim,Shuzhen Hong,et al. Determination of DNA Binding Behavior of FoxA1 Constructs Using a Gold Nanoparticle-Based High Throughput Assay[J]. journal of Molecular and Engineering Materials,2017,4(3).
APA Khin Moh Moh Aung,Michelle Gek Liang Lim,Shuzhen Hong,Edwin Cheung,&Xiaodi Su.(2017).Determination of DNA Binding Behavior of FoxA1 Constructs Using a Gold Nanoparticle-Based High Throughput Assay.journal of Molecular and Engineering Materials,4(3).
MLA Khin Moh Moh Aung,et al."Determination of DNA Binding Behavior of FoxA1 Constructs Using a Gold Nanoparticle-Based High Throughput Assay".journal of Molecular and Engineering Materials 4.3(2017).
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