Establishment of estrogen receptor α trans-activation system
- VernacularTitle:雌激素受体α转录激活系统的构建
- Author:
Jieping LI
;
Zhihong XIONG
;
Zhihong YANG
;
Zhaoyun WANG
;
Liping YANG
;
Qinong YE
- Publication Type:Journal Article
- Keywords:
Receptors,estrogen;
Transcription,genetic;
Transfection;
Polymerase chain reaction
- From:
Chinese Journal of Obstetrics and Gynecology
2008;43(8):611-614
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct estrogen receptor α (ERα)trans-activation system. Methods The full length ERα and its different function regions [ ( transcriptional activation function 1 ( AF1 ), DNA inding domain ( DBD), and transcriptional activation function 2 ( AF2 ) ] were amplified from pcDNA3 -ERα by PCR and cloned into the pGAL vector. The expressions of the recombinant plasmids constructed were detected via immunoblotting. The 293T cells transfected with recombinant plasmids of full length ERα, its different function regions and empty vector were divided into 5 groups; each group was divided into 2 parts which were treated with or without estrogen (E2). The transcriptional activity of each group was detected in 293T cells after the recombinant plasmid was co-transfected with 0. 2 μg of estrogen receptor element luciferase(ERE-LUC) and 0. 1 μg of plasmid expressing β-galactosidase and treated with or without 10 nmol/L E2 for 24 hours. Results The full length ERα and its different function regions were expressed in the 293T cells. Compared with the empty pGAL vector, the transcription activities of full length ERα, AF1, AF2 and DBD recombinant plasmids were raised about 20. 44±1.01, 2. 09±0. 11, 8. 09±0. 30 and 1.05±0. 09 fold, respectively, with the induction of E2 after transfection in the 293T colls. Conclusion The trans-activation system of ERα has been successfully established.
