Τίτλος:
Application of denaturing gradient gel electrophoresis (DGGE) to screen
for mutations of the human glucocorticoid receptor a gene (hGR alpha)
Γλώσσες Τεκμηρίου:
Αγγλικά
Περίληψη:
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.
Συγγραφείς:
Tsolakidou, AF
Coulocheri, SA
Sekeris, CE
Moutsatsou, P
Περιοδικό:
Clinical Biochemistry
Εκδότης:
PERGAMON-ELSEVIER SCIENCE LTD
Λέξεις-κλειδιά:
glucocorticoid receptor alpha; gene; mutation; denaturing-gradient gel
electrophoresis; glucocorticoid resistance; screening
DOI:
10.1016/S0009-9120(03)00027-4
