Investigations of glucocorticoid receptor mutant without ligand binding domain
- VernacularTitle:缺失激素结合区的人糖皮质激素受体突变体的研究
- Author:
Fang ZHANG
- Publication Type:Journal Article
- Keywords:
Glucocorticoid receptor;
Ligand binding domain;
Transcriptional activation activity
- From:
Journal of Medical Postgraduates
2003;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct expression vector of human glucocorticoid receptor mutant.Methods:cDNA sequence of human glucocorticoid receptor mutant having no LBD was amplificated through RT-nested PCR,PCR product was directionally cloned to pEGFP-N2 plasmid vector,and recombinant plasmids were screened through sequencing.Expressions of recombinant plasmids in human macrophage cell line U937 were observed through fluorescence microscope.Transcriptional activation activity of variant was evaluated through methods of relative activity of luciferase.Results:The 1 601 bp length cDNA was obtained,and result of sequencing confirmed the cloned cDNA sequence is completely coincidence with that of glucocorticoid receptor in Genebank.Expression products of recombinant plasmid mainly located in nucleus.Transcriptional activation activity incresed after recombinant plasmid transfected.Conclusion:Expression vector of glucocorticoid receptor mutant was constructed successfully,which has transcriptional activation activity independent of glucocorticoid.
