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Molecular organization of the human Raf-1 promoter region.

机译:人类Raf-1启动子区域的分子组织。

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摘要

A genomic DNA fragment containing the Raf-1 promoter region was isolated by using a cDNA extension clone. Nucleotide sequencing of genomic DNA clones, primer extension, and S1 nuclease assays have been used to identify the 5' ends of Raf-1 RNAs. Consistent with its ubiquitous expression, the Raf-1 promoter region had features of a housekeeping gene in that it was GC-rich (HTF-like), lacked TATA and CAAT boxes, and contained heterogeneous RNA start sites and four potential binding sites for the transcription factor SP1. In addition, an octamer motif (ATTTCAT), a potential binding site for the octamer family of transcription factors, was located at -734 base pairs. The Raf-1 promoter region drove reporter gene expression 30-fold over the promoterless reporter in Cos 7 cells.
机译:通过使用cDNA延伸克隆分离含有Raf-1启动子区域的基因组DNA片段。基因组DNA克隆的核苷酸测序,引物延伸和S1核酸酶检测已用于鉴定Raf-1 RNA的5'末端。与其普遍表达一致,Raf-1启动子区域具有管家基因的特征,因为该区域富含GC(类似于HTF),缺少TATA和CAAT框,并且包含异源RNA起始位点和四个潜在的结合位点。转录因子SP1。此外,八聚体基序(ATTTCAT)是转录因子八聚体家族的潜在结合位点,位于-734个碱基对处。 Raf-1启动子区域驱动报告基因在Cos 7细胞中的表达比无启动子报告基因高30倍。

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