Puma luciferase reporter gene construction and identification
10.3760/cma.j.issn.1006-9801.2011.01.003
- VernacularTitle:Puma荧光素酶报告基因的构建和鉴定
- Author:
Xin YANG
;
Shi QIU
;
Shouzhi GU
;
Yun CAI
;
Xing GAO
;
Zejun LIU
- Publication Type:Journal Article
- Keywords:
Puma;
Promoter;
Luciferase;
Reporter gene;
Apoptosis
- From:
Cancer Research and Clinic
2011;23(1):8-10
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the mechanism of p55 inducing cell apoptosis, the 180 bp fragment of Puma promoter was cloned into the pGL3-basic luciferase reporter vector. The biological activity of Pumareporter plasmid was verified by cell transfection. Methods The target fragments of Puma were amplified by RT-PCR method and the fragments were inserted into the pGL3-basic luciferase reporter vector. The acquired Puma-Luc plasmid was transfected into H1299 cell line and detected its activity. Results Sequencing indicated that the amplified Puma promoter is correct. Dual-luciferase Reporter Assay showed the Puma-Luc constructs have promoter activity. Conclusion The cloning of human Puma gene promoter and the construction of its reporter vector were successful. This study will lay the foundation for further research on the function of p53 inducing apoptosis through mitochondrial pathway.
