秦川牛PLAG1基因启动子克隆及转录因子预测

doi:10.16431/j.cnki.1671-7236.2023.07.007

摘要/Abstract

摘要： 【目的】探究秦川牛多形性腺瘤基因1（pleomorphic adenoma gene 1，PLAG1）的组织表达规律，并克隆其启动子区序列，预测分析其关键转录因子结合位点，为探究其转录调控机制提供理论参考。【方法】采集3头20月龄秦川牛成年公牛心脏、肝脏、脾脏、肺脏、肾脏、皮下脂肪、背最长肌、瘤胃组织，用实时荧光定量PCR方法检测PLAG1基因在不同组织中的相对表达量，同时克隆PLAG1基因上游启动子区序列，利用生物信息学软件预测PLAG1基因转录起始位点及启动子核心区域，分析、筛选核心启动子区域的关键转录因子结合位点。【结果】PLAG1基因在秦川牛各组织中均有表达，且在背最长肌中的表达量显著高于其他组织（P<0.05）。PLAG1基因启动子序列全长1 861 bp，生物信息学预测分析发现PLAG1基因核心启动子区位于-297―＋42 bp，存在高度保守的Krüppel样因子5（KLF5）、cAMP反应元件结合蛋白1（CREB1）和早期生长反应因子1（EGR1）转录因子结合位点，且CpG岛位于PLAG1基因核心启动子区域内。【结论】PLAG1基因在肌肉组织中高表达，其核心启动子区位于-297―＋42 bp，KLF5、CREB1和EGR1转录因子对PLAG1基因的转录活性可能具有调控作用。本研究结果为进一步探究PLAG1基因的转录调控机制提供了理论依据。

关键词: 秦川牛; PLAG1基因; 启动子; 转录因子

Abstract: 【Objective】 The purpose of this study was to explore the histological expression rules of pleomorphic adenoma gene 1 (PLAG1) in Qinchuan cattle, clone the promoter sequences of PLAG1 gene, predict and analyze the key transcription factor binding sites, so as to provide theoretical reference for exploring the transcriptional regulation mechanism of PLAG1 gene.【Method】 The relative expression of PLAG1 gene in the heart, liver, spleen, lung, kidney, subcutaneous fat, longissimus dorsi muscle and rumen of three 20-month-old Qinchuan adult bulls were detected by Real-time quantitative PCR, and the sequences of the upstream promoter region of PLAG1 gene were cloned.Bioinformatics software was used to predict PLAG1 gene transcription start sites and promoter core regions, and to analyze and screen key transcription factor binding sites in core promoter regions.【Result】 PLAG1 gene was expressed in all tissues of Qinchuan cattle, and the expression of PLAG1 gene in longissimus dorsi muscle was significantly higher than that in other tissues (P<0.05).The full length of the PLAG1 gene promoter sequence was 1 861 bp, and bioinformatics prediction analysis found that the core promoter region of the PLAG1 gene was located at -297-+42 bp. There were highly conserved transcription factor binding sites for Krüppel-like factor 5 (KLF5), cAMP-responsive element-binding protein-1 (CREB1) and early growth response factor 1 (EGR1), and the CpG island was located in the core promoter region of PLAG1 gene.【Conclusion】 PLAG1 gene was highly expressed in muscle tissue, and its core promoter region was located at -297-+42 bp.Transcription factors KLF5, CREB1 and EGR1 might have a regulatory effect on transcriptional activity of PLAG1 gene.The results provided theoretical basis for further investigation of PLAG1 gene transcriptional regulation mechanism.

Key words: Qinchuan cattle; PLAG1 gene; promoter; transcription factor

中图分类号:

S823

马勇, 陈宗昌, 李旭鹏, 李静生, 白彦斌, 刘占鑫, 魏亚丽, 郭达山, 石斌刚, 赵志东, 罗玉柱, 胡江, 王继卿, 刘秀, 李少斌. 秦川牛PLAG1基因启动子克隆及转录因子预测[J]. 中国畜牧兽医, 2023, 50(7): 2661-2669.

MA Yong, CHEN Zongchang, LI Xupeng, LI Jingsheng, BAI Yanbin, LIU Zhanxin, WEI Yali, GUO Dashan, SHI Bingang, ZHAO Zhidong, LUO Yuzhu, HU Jiang, WANG Jiqing, LIU Xiu, LI Shaobin. Cloning and Transcription Factor Prediction of PLAG1 Gene in Qinchuan Cattle[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(7): 2661-2669.

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