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PRR11(proline-rich protein 11,PRR11)是我们最近发现的一个新的肿瘤相关基因.初步研究表明,PRR11参与细胞增殖、细胞周期和细胞癌变等多种生物学过程.为了进一步研究PRR11基因的转录调控机制并全面解析其功能,本研究对PRR11基因的启动子进行了克隆鉴定和初步分析.首先,应用5’RACE(rapid amplification of cDNA ends,cDNA末端快速扩增)技术鉴定了PRR11基因的转录起始位点,发现了其具有多个转录起始位点.通过PCR定向克隆和DNA blunting技术,构建了6个相互重叠并覆盖PRR11基因转录起始位点附近约2.0 kb区域的PRR 11基因启动子荧光素酶报告基因重组体.启动子活性分析表明,PRR 11基因启动子主要定位于转录起始位点附近-563 bp～+341 bp的区域内.采用转录因子结合位点预测分析软件分析表明,PRR 11基因启动子缺乏典型的TATA盒,但含有典型的GC盒、CCAAT盒以及潜在的经典转录因子E2F1和MYB的结合位点,提示Sp1、NF-Y、E2F1和MYB等经典转录因子可能参与PRR 11基因的转录调控.
PRR11 (PRR11) is a new tumor-associated gene that we recently discovered.Preliminary studies show that PRR11 is involved in many biological processes such as cell proliferation, cell cycle and cell carcinogenesis.In order to further study the transcription of PRR11 gene Regulatory mechanism and a comprehensive analysis of its function, the PRR11 gene promoter cloning identification and preliminary analysis.First, the use of 5’RACE (rapid amplification of cDNA ends, rapid amplification of cDNA ends identified PRR11 gene transcription And found that it has multiple transcriptional start sites.Through PCR directed cloning and DNA blunting technology, six PRR 11 genes overlapping each other and covering about 2.0 kb near the transcription initiation site of PRR11 gene were constructed Promoter luciferase reporter gene recombinants.The promoter activity analysis showed that the PRR 11 gene promoter mainly located in the region of -563 bp to + 341 bp near the transcription start site.The transcription factor binding site prediction analysis software The analysis showed that the PRR 11 gene promoter lacks the typical TATA box but contains the combination of the typical GC box, CCAAT box and the potential classical transcription factors E2F1 and MYB Point, suggesting Sp1, NF-Y, E2F1 and MYB transcription factors may be involved in other classical transcriptional regulation of PRR 11 gene.