Authors: Ronai Z.; Guttman A.; Keszler G.; Sasvari-Szekely M.

Source: Current Medicinal Chemistry, Volume 11, Number 8, April 2004 , pp. 1023-1029(7)

Publisher: Bentham Science Publishers

Abstract:

DNA-protein interaction in the 5' upstream polymorphic region of the dopamine D4 receptor (DRD4) gene was analyzed by capillary electrophoretic mobility shift assay (CEMSA). The sequence of interest was amplified using a fluorescent primer and applied as a probe in the binding assays with HeLa nuclear extract. Serial dilution of the probe resulted in a concentration dependent DNA-protein complex formation. Sp 1 specific oligonucleotide competitor significantly inhibited the DNA-protein complex formation. A non-specific competitor, differing only in three base pairs, showed weaker effect pointing to the contribution of the Sp 1 recognition sequence in the complex. Polymorphic competitors were also prepared from homozygous individuals possessing either duplicated (2x120 bp) or single copy (1x120bp) of the 120 bp repeat sequence and were used against the Sp 1 specific probe in competition assays. Our data provide experimental evidence for the binding of Sp 1 to the 120 bp duplicated sequence of the DRD4 5' upstream region and suggest enhanced binding capacity of the duplicated form.

Keywords: capillary electrophoresis; electrophoretic mobility shift assay; dopamine d4 receptor

Document Type: Review article

DOI: http://dx.doi.org/10.2174/0929867043455503

Affiliations: 1: Institute of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, H-1088 Budapest, Puskin u.9., Hungary.

Publication date: 2004-04-01

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• Current Medicinal Chemistry covers all the latest and outstanding developments in medicinal chemistry and rational drug design. Each issue contains a series of timely in-depth reviews written by leaders in the field covering a range of the current topics in medicinal chemistry. Current Medicinal Chemistry is an essential journal for every medicinal chemist who wishes to be kept informed and up-to-date with the latest and most important developments.

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