Application of denaturing gradient gel electrophoresis (DGGE) to screen for mutations of the human glucocorticoid receptor alpha gene (hGRalpha)
| Autor: | Constantine E. Sekeris, Stavroula A. Coulocheri, Amalia F. Tsolakidou, Paraskevi Moutsatsou |
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| Rok vydání: | 2003 |
| Předmět: |
chemistry.chemical_classification
Genetics Mutation DNA Complementary Clinical Biochemistry Mutant DNA Mutational Analysis Wild type Reproducibility of Results General Medicine DNA-binding domain Biology medicine.disease_cause Polymerase Chain Reaction Sensitivity and Specificity Glucocorticoid receptor Receptors Glucocorticoid chemistry Complementary DNA medicine Humans Nucleotide Electrophoresis Polyacrylamide Gel Gene |
| Zdroj: | Clinical biochemistry. 36(4) |
| ISSN: | 0009-9120 |
| Popis: | Objectives In a previous publication, we had presented a sensitive method to detect mutations of the segment of the human glucocorticoid receptor alpha (hGRα) gene encoding the ligand binding domain (LBD) and part of the DNA binding domain (DBD) of hGRα, 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 hGRα are increasingly reported in a variety of disease states. We now present an expanded screening methodology to detect mutations covering the whole length of hGRα. 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 hGRα. Wild type hGRα 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 hGRα. 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 hGRα mutations in the aetiopathology of diseases correlated to derangements of glucocorticoid action. |
| Databáze: | OpenAIRE |
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