Transcriptional regulation of GCLC gene in human bronchus epithelial cells
- VernacularTitle:人支气管上皮细胞?-谷氨酰半胱氨酸合酶催化亚单位基因转录的调控
- Author:
Guoming ZOU
;
Bing LI
;
Pixin RAN
- Publication Type:Journal Article
- Keywords:
Genes;
Transcriptional regulation;
Glutathione
- From:
Chinese Journal of Pathophysiology
2000;0(08):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To investigate the transcriptional regulating function of the catalytic subunit(GCLC)gene of ?-glutamylcysteine synthase(?-GCS),the rate-limiting enzyme of glutathione(GSH)synthetic enzymes.METHODS:The regulating region of the human GCLC gene was cloned by PCR method to construct the wild type plasmid,which expressed luciferase reporting gene.Many mutated plasmids expressing luciferase reporting gene were also constructed by the method of exonuclease Ⅲ with S1 nuclease.The transcription regulation of GCLC gene was investigated by transient transfection of the wild type and the mutated plasmids through the observation of relative activities of luciferase.RESULTS:We found that the-2 515～-2 236 bp,-864～-838 bp,-769～-538 bp,-421～-341 bp regions upside from the transcriptional start site up-regulated the gene transcription are the new positive transcription-regulating regions and the-810～-769 bp(especial the-782～-769 bp)region is the negative transcription-regulating region,which is more precise than ever.CONCLUSION:The results of this experiment provide one good foundation to study the transcription management of the GCLC gene and GSH synthesis.
