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Title 

Directed immobilization of DNA-binding proteins on a cognate DNA-modified chip surface

Authors 

Eun Ju JeongYoo Seok JeongKyoungsook ParkSo Yeon YiJun Hyoung AhnSang Jeon ChungMoonil KimBong Hyun Chung

Publisher 

Elsevier

Issue Date 

2008

Citation 

Journal of Biotechnology, vol. 135, no. 1, pp. 16-21

Keywords 

DNA-binding domainDNA-modified surfacesite-directed immobilizationSPR imagingbiochipsDNAimaging systemssurface plasmon resonancetranscription factorsbinding profiling

Abstract 

Here we describe a useful method for the site-directed immobilization of proteins with a DNA-binding domain (DNA-BD) on the cognate DNA-coated gold surface for surface plasmon resonance (SPR) imaging analyses. In order to assess the performance of this procedure, we utilized two DNA-BDs, yeast GAL4 DNA-BD, and bacterial LexA DNA-BD. After the immobilization of the cognate double-stranded DNAs (dsDNAs) to a gold chip surface with a monolayer of poly(l-lysine) for sequence-specific DNA-protein interaction, purified recombinant GAL4 DNA-BD:EGFP and LexA DNA-BD:RFP fusion proteins were applied to a dsDNA-spotted gold chip, and were subsequently analyzed using an SPR imaging system. Consequently, the recombinant DNA-binding proteins, GAL4 DNA-BD:EGFP and LexA DNA-BD:RFP, were shown to bind selectively to their cognate DNA sequences on the gold chip. Collectively, our results revealed that sequence-specific dsDNA microarray approach could prove useful in performing the site-directed immobilization of DNA-binding proteins onto a gold thin film in a parallel format, and thereby potentially allowing for the analysis of transcription factor binding profiling as well as for the monitoring of protein-protein interactions between target proteins with DNA-binding domain as a fusion tag and their binding partners.

ISSN 

0168-1656

Link 

http://dx.doi.org/10.1016/j.jbiotec.2008.02.019

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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