Establishment of chromatin immunoprecipitation assay to analyze transcriptional regulation of gene expression
- VernacularTitle:运用染色质免疫沉淀技术分析基因表达转录调控方法的建立及初步应用
- Author:
Liping ZHANG
- Publication Type:Journal Article
- Keywords:
Chromatin immunoprecipitation assay;
Transcriptional regulation;
PEPCK
- From:
Chinese Journal of Laboratory Medicine
2001;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a Chromatin immunoprecipitation assay to analyze transcriptional regulation of gene expression in hepatocytes. Methods The PEPCK gene was used as a target gene, HPRT, MBP and IgG were used as internal reference, the chromatin immunoprecipitation and PCR were performed with antibody specific for CREB to verify bindings of transcription factor CREB to the PEPCK promoter under various metabolic conditions. Results Transcription factor CREB occupies its target sites in the PEPCK promoter and this binding is increased by 4-fold during fasting in the anti-CREB antibody immunoprecipitated DNA fragments. Conclusion The results confirm that CREB binds to the PEPCK promoter and is involved in transcriptional regulation of PEPCK gene in hepatocytes.
