| 引用本文: | 宋崇阳,张思勉,李永闯,张攀,赖晓芳,王攀攀,于飞,高焕,阎斌伦.脊尾白虾Dnmt2启动子克隆及其转录调控分析[J].海洋科学,2022,46(5):95-103. |
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| 脊尾白虾Dnmt2启动子克隆及其转录调控分析 |
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宋崇阳1, 张思勉1, 李永闯1, 张攀1, 赖晓芳1, 王攀攀1,2,3, 于飞4, 高焕1,2,3, 阎斌伦1,2,3
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1.江苏海洋大学 江苏省海洋生物资源与环境重点实验室 江苏省海洋生物技术重点实验室, 江苏 连云港 222005;2.江苏省海洋生物产业技术协同创新中心, 江苏 连云港 222005;3.江苏省农业种质资源保护与利用平台, 江苏 南京 210014;4.连云港市海洋与渔业发展促进中心, 江苏 连云港 222005
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| 摘要: |
| 为探明脊尾白虾(Exopalaemon carinicauda) Dnmt2 (DNA methyltransf-erase2, Dnmt2)启动子对Dnmt2转录的调控作用, 本研究利用染色体步移技术克隆得到脊尾白虾Dnmt2启动子, 使用在线软件预测该基因启动子区域的转录调控元件及转录因子结合位点, 通过构建一系列缺失表达载体, 利用双荧光素酶报告基因检测技术鉴定了该基因核心启动子区, 并对其启动子活性进行检测。结果表明, 克隆的脊尾白虾Dnmt2启动子长度为1315 bp, 转录起始位点“A”位于ATG上游236 bp, 启动子区域含有多种转录因子结合位点, 包括STAT3、SOX、NF-κB、E2F、GATA-1等, 但缺乏CpG岛。双荧光素酶报告基因检测显示, Dnmt2核心启动子区位于–196~+81 bp, 对启动子区域进行活性检测发现, 在该基因启动子–640~–452 bp区域内可能存在促进基因表达的转录调控元件, 而在–1160~ –640 bp区域可能存在抑制该基因表达的转录调控元件。本研究结果为进一步研究Dnmt2启动子的表达调控机制提供理论依据。 |
| 关键词: 脊尾白虾(Exopalaemon carinicauda) Dnmt2 核心启动子 启动子活性 转录调控元件 |
| DOI：10.11759/hykx20211207001 |
| 分类号:Q785;S917.4 |
| 基金项目:江苏海洋大学江苏省海洋生物资源与环境重点实验室开放课题基金资助(SH20191203); 江苏省优势学科建设工程资助项目(PAPD); 江苏省研究生科研与实践创新计划项目(KYCX20-2877); 江苏海洋大学大学生创新创业训练计划项目(SZ202111641631003) |
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| Cloning and transcriptional regulation of Dnmt2 promoter in Exopalaemon carinicauda |
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SONG Chong-yang1, ZHANG Si-mian1, LI Yong-chuang1, ZHANG Pan1, LAI Xiao-fang1, WANG Pan-pan1,2,3, YU Fei4, GAO Huan1,2,3, YAN Bin-lun1,2,3
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1.Jiangsu Ocean University, Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Key lab of Marine Biotechnology, Lianyungang 222005, China;2.Co-Innovation Center of Jiangsu Marine Bio-industry Technology, Lianyungang 222005, China;3.The Jiangsu Provincial Infrastructure for Conservation and Utilization of Agricultural Germplasm, Nanjing 210014, China;4.Lianyungang Marine and Fishery Development Promotion Center, Lianyungang 222005, China
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| Abstract: |
| In order to explore the regulation of Dnmt2 promoter of Exopalaemon carinicauda on Dnmt2 transcription, the Dnmt2 promoter of E. carinicauda was cloned by chromosome walking technique, and the transcription regulatory elements and transcription factor binding sites in the promoter region of the gene were predicted by online software, By constructing a series of deletion expression vectors, the core promoter region of the gene was identified by double luciferase reporter gene detection technology, and its promoter activity was detected. The results showed that the length of the cloned Dnmt2 promoter was 1315 bp, the transcription start site "A" was 236 bp upstream of ATG, and the promoter region contained a variety of transcription factor binding sites, including STAT3, Sox, NF- κ B, E2F, GATA-1, etc., but lacking CpG island. The detection of double luciferase reporter gene showed that the core promoter region of Dnmt2 was located at –196~+81 bp. The activity detection of the promoter region showed that there may be transcriptional regulatory elements promoting gene expression in the –640~–452 bp region of the gene promoter, while there may be transcriptional regulatory elements inhibiting gene expression in the –1160~–640 bp region. The results of this study provide a theoretical basis for further study on the expression regulation mechanism of Dnmt2 promoter. |
| Key words: Exopalaemon carinicauda Dnmt2 core promoter promoter activity transcriptional regulatory element |
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