Beck T W, Brennscheidt U, Sithanandam G, Cleveland J, Rapp U R
Program Resources, Inc., Biological Carcinogenesis Development Program, Frederick, Maryland 21701.
Mol Cell Biol. 1990 Jul;10(7):3325-33. doi: 10.1128/mcb.10.7.3325-3333.1990.
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.
利用cDNA延伸克隆分离出一个包含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核酸酶分析已用于鉴定Raf-1 RNA的5'末端。与其广泛表达一致,Raf-1启动子区域具有管家基因的特征,即富含GC(类HTF),缺乏TATA盒和CAAT盒,包含异质性RNA起始位点以及转录因子SP1的四个潜在结合位点。此外,一个八聚体基序(ATTTCAT),即转录因子八聚体家族的潜在结合位点,位于-734碱基对处。在Cos 7细胞中,Raf-1启动子区域驱动报告基因的表达比无启动子的报告基因高30倍。