Construction of series reporter plasmids with truncated and deleted human telomerase reverse transcriptase promoter
10.3760/cma.j.issn.0254-5101.2012.04.004
- VernacularTitle:系列截短和缺失的hTERT启动子报告载体的构建及活性验证
- Author:
Fang LUAN
;
Hua LIU
;
Bin LIU
;
Chunhong MA
- Publication Type:Journal Article
- Keywords:
Human telomerase reverse transcriptase;
Reporter plasmid;
Promoter activity
- From:
Chinese Journal of Microbiology and Immunology
2012;32(4):309-314
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct series of reporter plasmids with truncated and deleted hTERT promoter.Methods Gene fragments of hTERT promoter was amplified by PCR and cloned into pGL3-Basic to construct luciferase reporter vectors.Dual luciferase assays were performed with cell lysates of HepG2 and COS-7 cells cotransfected with hTERT promoter reporter plasmids and pRL-TK.Results Series of luciferase reporter plasmids with truncated and deleted hTERT promoter were successfully constructed and respectively named pGL3B-895,pGL3B-371,pGL3B-DELS2,pGL3B-349,pGL3B-329,pGL3B-318,pGL3B-306.Dual luciferase reporter assays showed that all the reporter vectors have promoter activity both in HepG2 and COS-7.Conclusion Series of luciferase reporter plasmids with truncated and deleted hTERT promoter were successfully constructed,and their promoter activity were verified.These plasmids provide necessary experimental naterials for further investigation of regulation of hTERT during hepatocarcinoma development.
