Effect of HBx on transcriptional activity of IGF-II gene P4 promoter
- VernacularTitle:HBx蛋白对IGF-II基因P4启动子活性的影响
- Author:
Shaohui TANG
;
Chunhui QU
;
Minying YANG
;
Yuhong LUO
;
Donghua YANG
- Publication Type:Journal Article
- Keywords:
Hepatitis B virus X protein;
P4 promoter;
Transcriptional activity
- From:
Chinese Journal of Pathophysiology
2010;26(1):7-11
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To construct HBx eukaryotic expression vector pEYFP-C1-X and eukaryotic expression vector pGL3-P4 driven by P4 promoter of human IGF-II gene and to investigate the effect of HBx on the transcription activity of IGF-II gene P4 promoter. METHODS: HBx gene and P4 promoters were cloned into pEYFP-C1 and pGL3-basic vectors respectively by gene recombination techniques to construct recombinant plasmids pEYFP-C1-X and pGL3-P4. HepG2 cells were transfected with pEYFP-C1-X and the resistant cell clones were selected by G418. Then methylated pGL3-P4 was transiently transfected into the above cell clones, and the transcription activity of P4 promoter was determined by dual-luciferase reporter assay system. RESULTS: (1) Aim fragments HBx gene and P4 promoter that were cloned were 465 bp and 1 246 bp, respectively and the DNA sequences were accordant with GenBank data confirmed by restricted enzyme digestion and sequencing. (2) HepG2-EYFP-X cells that expressed HBx protein were obtained. (3) Luciferase activity of methylated P4 promoter in the HepG2-EYFP-X was more than that of control cell HepG2-EYFP (P<0.01). CONCLUSION: HBx may enhance the transcription activity of the P4 promoter.
