Characterization of Porcine Nanog 5’-flanking Region

초록

Nanog is a homeodomain proteins and functions self-proliferation of embryonic stem cells to maintain the pluripotency. There are several reports regarding the function of human and mouse Nanog gene except pig. Therefore, in this study, we are trying to uncover the porcine Nanog basal promoter activity that is applicable of possible ES cells marker in pig. To characterize the porcine Nanog promoter, 5'-flanking region of Nanog was isolated from the cells of the Mini-pig ears. Deletion fragments from the 5'-flanking region of the relative activity was measured using the Dual-Luciferase Reporter Assay System, and a fluorescence marker, GFP. We found that there are basal regulatory factors from —99 to +194 for expression of porcine Nanog gene using deletion fragment activity. We thought Sp1 transcription binding site as a possible regulatory transcriptional site and confirmed its activity with deletion mutant fragment. Gel shift and CHIP analysis showed that Sp1 protein bind to the site we checked. These results show that Sp1 transcription factor is essential element for porcine Nanog basal activity as it works in human and mouse.