Application of denaturing gradient gel electrophoresis (DGGE) to screen for mutations of the human glucocorticoid receptor a gene (hGR alpha)

Tsolakidou, AF; Coulocheri, SA; Sekeris, CE; Moutsatsou, P

dc.contributor.author	Tsolakidou, AF	en
dc.contributor.author	Coulocheri, SA	en
dc.contributor.author	Sekeris, CE	en
dc.contributor.author	Moutsatsou, P	en
dc.date.accessioned	2014-03-01T01:53:05Z
dc.date.available	2014-03-01T01:53:05Z
dc.date.issued	2003	en
dc.identifier.issn	0009-9120	en
dc.identifier.uri	https://dspace.lib.ntua.gr/xmlui/handle/123456789/26844
dc.subject	glucocorticoid receptor alpha	en
dc.subject	gene	en
dc.subject	mutation	en
dc.subject	denaturing-gradient gel electrophoresis	en
dc.subject	glucocorticoid resistance	en
dc.subject	screening	en
dc.subject.classification	Medical Laboratory Technology	en
dc.subject.other	HORMONE-BINDING DOMAIN	en
dc.subject.other	SHIFT MUTATION	en
dc.subject.other	IN-VIVO	en
dc.subject.other	RESISTANCE	en
dc.subject.other	ASSOCIATION	en
dc.subject.other	VARIANT	en
dc.subject.other	BETA	en
dc.subject.other	GR	en
dc.subject.other	POLYMORPHISM	en
dc.subject.other	SENSITIVITY	en
dc.title	Application of denaturing gradient gel electrophoresis (DGGE) to screen for mutations of the human glucocorticoid receptor a gene (hGR alpha)	en
heal.type	journalArticle	en
heal.language	English	en
heal.publicationDate	2003	en
heal.abstract	Objectives: In a previous publication, we had presented a sensitive method to detect mutations of the segment of the human glucocorticoid. receptor alpha (hGRalpha) gene encoding the ligand binding domain (LBD) and part of the DNA binding domain (DBD) of hGRalpha, as several types of glucocorticoid resistance syndromes have been correlated with mutations in the respective nucleotide sequences. However, mutations affecting various regions covering the whole length of hGRalpha are increasingly reported in a variety of disease states. We now present an expanded screening methodology to detect mutations covering the whole length of hGRa. Design and Methods: We developed a sensitive, simple screening PCR-DGGE method to detect mutations in the aminoterminal domain and DNA-binding domain of the hGRa. Wild type hGRa cDNA and mutant samples were included in the analysis to ensure the accuracy and sensitivity of the method. Results: The PCR-DGGE method identified the mutant samples and discriminated them from wild type hGRa. Conclusions: The method described is accurate, sensitive, simple, cheap and fulfills the critera for a screening method which will be useful in delineating possible involvement of hGRalpha mutations in the aetiopathology of diseases correlated to derangements of glucocorticoid action. (C) 2003 The Canadian Society of Clinical Chemists. All rights reserved.	en
heal.publisher	PERGAMON-ELSEVIER SCIENCE LTD	en
heal.journalName	CLINICAL BIOCHEMISTRY	en
dc.identifier.isi	ISI:000183789000011	en
dc.identifier.volume	36	en
dc.identifier.issue	4	en
dc.identifier.spage	305	en
dc.identifier.epage	311	en