| 徐苙菲,刘阳嘉,庄小琼,谢莉萍,张荣庆.合浦珠母贝SOX9对Prismalin-14基因的调控研究[J].现代生物医学进展英文版,2018,(1):1-5. |
| 合浦珠母贝SOX9对Prismalin-14基因的调控研究 |
| Research on the Regulation of Prismalin-14 gene by SOX9 Protein in Pinctada Fucata |
| Received:May 15, 2017 Revised:June 08, 2017 |
| DOI：10.13241/j.cnki.pmb.2018.01.001 |
| 中文关键词: 合浦珠母贝 SOX9 Prismalin-14 转录调控 |
| 英文关键词: Pinctada Fucata SOX9 Prismalin-14 Transcription Regulation |
| 基金项目:国家自然科学基金面上项目（31372508） |
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| 中文摘要: |
| 摘要 目的：研究合浦珠母贝转录因子SOX9对Prismalin-14的转录调控机制。方法：应用在线预测软件PROMO分析Prismalin-14的启动子序列，以预测Prismalin-14启动子上可能的转录因子与其结合位点；运用细胞共转染实验和双荧光素酶报告系统以检测SOX9对Prismalin-14启动子的激活作用；构建Prismalin-14启动子截短体的荧光素酶报告载体，并和SOX9的真核载体共转到HEK-293T细胞中，再进行双荧光素酶报告系统检测Prismalin-14启动子的活性；构建SOX9截短体的真核表达载体，并与Prismalin-14启动子的荧光素酶报告载体共转到HEK-293T细胞中，再进行双荧光素酶报告分析Prismalin-14启动子的活性。结果：SOX9能激活Prismalin-14的启动子的活性，并具有剂量效应；对Prismalin-14启动子进行截短后，不包含结合位点的Prismalin-14启动子的活性是野生型Prismalin-14启动子活性的49 %，推测Prismalin-14启动子上的-415bp到-405bp区域是SOX9激活作用的关键区域；对SOX9的SRY-related HMG结构域进行截短后，其对Prismalin-14启动子的激活作用显著减少，因此SOX9结构的完整对Prismalin-14启动子活性的激活作用是必须的。结论：Prismalin-14的转录可能受SOX9调控，为进一步研究合浦珠母贝的转录调控机制提供基础，将有助于从分子水平上理解贝壳形成的上游调控机理。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the transcription regulation mechanism of Prismalin-14 by transcription factor SOX9 in the pearl oyster, Pinctada Fucata. Methods: The promoter sequence of Prismalin-14 was analyzed by using the online prediction software PROMO to predict the possible transcription factors on the Prismalin-14 promoter and its binding sites. Cell co-transfection experiments and dual luciferase reporter assay system were used to detect the activation of SOX9 on Prismalin-14 promoter. Constructed the luciferase assay vector of the truncated promoter of the Prismalin-14 and co-transfected into the HEK-293T cells with the eukaryotic ex- pression vector of the SOX9. Then, the dual luciferase reporter assay system was used to analyze the activity of the Prismalin-14 promoter. Constructed the eukaryotic expression vector of the truncated the SOX9 and co-transfected into the HEK-293T cells with the luciferase assay vector of the Prismalin-14. Then, dual luciferase reporter assay system was used to analyze the activity of the Prismalin-14 promoter. Results: SOX9 up-regulated the promoter activity of the Prismalin-14 with a dose effect. After truncated the Prismalin-14 promoter, the activity of the promoter of the Prismalin-14 without the binding site was 49 % of the Prismalin-14 promoter activity. As a result, the re- gion -415 bp to -405 bp of the Prismalin-14 promoter is the key region for activation activity of SOX9. After truncated the SRY-related HMG domain of SOX9, the activation of the Prismalin-14 promoter was significantly reduced. As a result, the structural integrity of SOX9 was essential for the promoter of the Prismalin-14 activation function. Conclusion: The transcription factor SOX9 regulates the expression of the Prismalin-14 in the pearl oyster, Pinctada fucata. It provided a basis for the further study of the transcription regulation mechanism in the pearl oyster, Pinctada fucata. It will help understand the upstream regulation mechanism of shell formation from the molecular level. |
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