猪TBX3启动子克隆和功能分析OA北大核心CSTPCD
Cloning and Functional Analysis of the Porcine T-box Transcription Factor 3 Promoter
猪(Sus scrofa)是重要的家畜,诱导干细胞的研究可以促进其育种行业的发展,T-Box转录因子3(T-box transcription factor 3,TBX3)对猪多能干细胞的干性具有重要的促进和维持作用.为了研究猪TBX3启动子的功能和调控机制,本研究通过序列比对和进化树构建对启动子区域保守性进行评估,并对启动子区域潜在的转录因子结合位点进行预测,对预测的转录因子进行GO和KEGG富集分析解释其参与的生物学过程,最终通过分子克隆和双荧光素酶报告系统验证启动子活性.结果显示,猪TBX3启动子预测区域在哺乳动物中共性程度较低,共性区域参与细胞增殖、分化、凋亡等过程;预测区域含有多种潜在的转录因子结合位点,其参与的信号通路包括干性维持和甲状腺激素信号通路;预测区域具有转录活性,其核心转录启动子处于-246/+64 bp的区域内.本研究通过探讨猪细胞中TBX3转录因子的上游调控机制,为深入理解猪TBX3的转录调控提供参考.
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.
程映皓;吴筱洁;沈巧艳;华进联
西北农林科技大学动物医学院/陕西省干细胞工程技术研究中心,杨凌 712100
畜牧业
T-Box转录因子3(TBX3)启动子转录因子预测双荧光素酶实验KEGG/GO富集分析
T-box transcription factor 3(TBX3)PromoterTranscription factor predictionDual-luciferase assayKEGG/GO enrichment analysis
《农业生物技术学报》 2024 (009)
2060-2070 / 11
国家重点研发计划(2022YFD1302201);国家自然科学基金(32072806;32372970);陕西省畜牧科技示范项目(20221086)
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