Related Experiment Videos
[A novel cis-acting enhancer element between CKLF and CKLFSF1 genes]
Ming-Xu Xu1, Mao-Yi Lai, Dong-Lan Xia
1Laboratory of Molecular Immunology, Human Disease Genomics, Peking University, Beijing 100083, China.
Aim:
To explore the roles of the CKLF gene and CKLFSF1 gene sequence (CCS) in transcriptional regulation.
Methods:
The target gene fragment was amplified by PCR and then inserted into pGL3-basic and pGL3-SV40 containing luciferase reporter vector gene to construct pGL3-basic-CCS and pGL3-SV40-CCS. Using liposome-mediated method, four recombinant plasmids were respectively transfected into Hela cells. Transient expression was analyzed.
Results:
The luciferase assay indicated that the no luciferase activity was detected in Hela cells transtected with pGL3-basic and pGL3-basic-CCS. However, the luciferase activity was doubled when pGL3-SV40-CCS was transfected into Hela cells.
Conclution:
The CCS has no promoter activity, whereas some important cis-acting enhancer elements which modulate its downstream gene expression may exist within this sequence.