Abstract As pigs (Sus scrofa) are important livestock animals, the study of porcine pluripotent induced stem cells can contribute to the development of the stockbreeding, T-box transcription factor 3 (TBX3) is important for promoting and maintaining stemness in porcine pluripotent stem cells. To investigate the function and regulatory mechanism of porcine TBX3 promoter, the conservativeness of the promoter region was assessed by sequence comparison and evolutionary tree construction, the potential transcription factor binding sites in the promoter region was also predicted and then the GO and KEGG enrichment analyses of the predicted transcription factors was performed to explain their involvement in the biological processes. Finally, the activity of promoter was verified by dual-luciferase reporter system. The results indicated that the predicted region of the porcine TBX3 promoter exhibited a low degree of commonality. The common region was found to be involved in cell proliferation, differentiation, apoptosis, and other processes. The predicted region was mined for a variety of potential transcription factors. The predicted region was confirmed to have binding sites and to participate in the signal pathway, including stemness maintenance and thyroid hormone signal. The predicted region was also confirmed to have transcriptional activity, with its core transcriptional promoter located within the region of -246/+64 bp. This study provides a reference for understanding the transcription regulatory mechanisms of TBX3 transcription factors in porcine cells.
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