Molecular cloning of HLA-B27 gene promoter and transcriptional activity detection in HeLa cells
- VernacularTitle:HLA-B27启动子荧光素酶报告基因载体的构建及其在Hela细胞的转录调控研究
- Author:
Li-Ke ZHAO
;
Jie-Ruo GU
;
Deen YU
;
- Publication Type:Journal Article
- Keywords:
HLA-B27 antigen;
Promoter regions(genetics);
Hela cell;
Activity regulation
- From:
Chinese Journal of Rheumatology
2003;0(08):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct pGL4.14-1uc eukaryotic expression vector for HLA-B27 promoter gene and explore the activity regulation of this promoter in Hela cells.Methods The HLA-B27 gene promot- er(-419 bp～1 bp)was amplified by polymerase chain reaction and was cloned into pGL4.14-luc vector to construct eukaryotic expression vector pGL4.14/B27 pro-luc.The purified pGL4.14/B27 pro-luc was stablely transfected into HeLa cells and the activity of HLA-B27 gene promoter was detected by luminometer.Results About 432 bp gene fragment was amplified by PCR from genomie DNA and pGL4.14/B27 pro-luc vector was constructed successfully.The activity of HLA-B27 promoter was 1.67?0.20,1.79?0.71,2.94?0.68,1.98?0.45 in Hela stable cells after treated with TNF-?,IFN-?,IFN-?and IFN-?for 48 hours.Conclusion TNF-?. IFN-?,IFN-?and IFN-?can regulate the B27 promoter activity.The high specific activity of constructed HLA-B27 promoter eukaryotic expression vector may be a good method for further research.
