杂色鲍HSP90基因启动子的功能分析

发布时间：2018-12-15 22:04

【摘要】：为探索启动子在热休克蛋白90表达调控中的作用,本研究在课题组已有杂色鲍HSP90基因cDNA的基础上,通过Genome walking、Tail-PCR和常规PCR等技术克隆获得该基因的5′调控区序列。在翻译起始位点(ATG)和第一外显子(长度94 bp)之间有一个809bp的内含子,第一外显子之前的5′调控区共2800 bp,从预测的转录起始位点(A)起,共2811 bp。在转录起始位点(A)上游 30 bp处存在TATA box。潜在的转录因子结合位点包括ATF、TBP、Sp1、Oct-1、C/EBPalpha、NF-1、NF-κappa B、GATA-1、Sox-2等。CpG岛预测软件分析其含1个CpG岛,长度为131 bp。实验构建了8个启动子缺失片段的萤火虫荧光素酶表达载体,通过瞬时转染293T细胞并进行双荧光素酶报告基因活性检测,确定杂色鲍HSP90基因核心启动子区位于 98~83 bp。在 624~ 539 bp,Oct-1、C/EBPalpha、NF-1这3个转录因子都起到一定的抑制作用。
[Abstract]:In order to explore the role of promoter in the regulation of heat shock protein 90 (HSP90) expression, the 5 'regulatory region of HSP90 gene was cloned by Genome walking,Tail-PCR and conventional PCR on the basis of cDNA of HSP90 gene. There is an intron of 809bp between the translation initiation site (ATG) and the first exon (94 bp in length), and the 5 '-regulatory region preceding the first exon 2800 bp, from the predicted transcriptional initiation site (A) to a total of 2811 bp.. TATA box. exists at 30 bp upstream of transcription initiation site (A) Potential transcriptional factor binding sites include ATF,TBP,Sp1,Oct-1,C/EBPalpha,NF-1,NF- 魏 appa, GATA-1, Sox-2 and so on. The CpG island prediction software analysis contains one CpG island and its length is 131 bp.. Eight fluorescent luciferase expression vectors with missing promoter fragments were constructed. By transient transfection of 293T cells and detection of the activity of double luciferase reporter gene, the core promoter region of HSP90 gene of abalone was determined to be located in the bp.. These three transcription factors all play a certain role in the inhibition of T624 ~ 539 bp,Oct-1,C/EBPalpha,NF-1.
【作者单位】：集美大学水产学院农业部东海海水健康养殖重点实验室福建;福建农林大学动物科学学院;
【基金】：国家自然科学基金(41176152) 集美大学创新团队基金(2010A001)~~
【分类号】：Q78;S917.4

【相似文献】