Cloning and analysis of promoter of pig copper zinc superoxide dismutase gene (CuZnSOD).
- Author:
Yuan SHI
;
Wei CHEN
;
Yongqing ZENG
;
Honglei ZHU
;
Zhenggang XU
;
Zhe ZHANG
;
Yun YANG
;
Tianyang ZHANG
- Publication Type:Journal Article
- MeSH:
Animals;
Cloning, Molecular;
Gene Expression Regulation;
Genetic Vectors;
Mice;
NIH 3T3 Cells;
Promoter Regions, Genetic;
Superoxide Dismutase;
genetics;
Swine;
Transfection
- From:
Chinese Journal of Biotechnology
2014;30(2):213-222
- CountryChina
- Language:Chinese
-
Abstract:
Pig copper zinc superoxide dismutase (CuZnSOD) is an important antioxidant enzyme. Some studies focused on the function of CuZnSOD gene, but the transcriptional regulation of the CuZnSOD gene is not yet fully elucidated. Therefore, the aims of the study were to determine the core promoter region and to explore its mechanism of transcriptional regulation. The 853 bp DNA sequence of 5'-flanking promoter was amplified by performing PCR. A series of CuZnSOD promoter fragments with gradually truncated 5'-end were produced by nested PCR and inserted into pGL3-Basic vector. The activities of the promoters were measured by the dual-luciferase assay system after transient transfection into the NIH/3T3 cells. The results demonstrated that there were 2 potential transcription start sites in the regions from initiation codon to -87 bp and -266 bp, respectively. The region from -383 bp to +67 bp in CuZnSOD gene promoter showed higher activity than other regions, and further deletion analysis demonstrated that the region from -75 bp to -32 bp contained an essential promoter sequence for pig CuZnSOD gene transcription. In addition, several potential transcription factor binding sites were predicted with bioinformatics method. These results suggest that these transcription factor binding sites may be involved in the transcriptional regulation of CuZnSOD gene.
