Falquerho L, Patey G, Paquereau L, Rossi V, Lahuna O, Szpirer J, Szpirer C, Levan G, Le Cam A
Centre CNRS-INSERM de Pharmacologie-Endocrinologie, Montpellier, France.
Gene. 1991 Feb 15;98(2):209-16. doi: 10.1016/0378-1119(91)90175-b.
The gene (PP63) encoding the inhibitor (PP63) of the insulin receptor tyrosine kinase was isolated from a rat genomic library. The intron/exon organization was deduced from Southern-blot analysis and sequence data (i.e., the exons + the boundaries). The PP63 gene, which maps to chromosome 11, spans approx. 8 kb and contains seven exons separated by six introns of different sizes. All of the boundaries match the consensus GT/AG sequence for donor and acceptor splice sites. Primer extension and S1 mapping experiments were used to locate the transcription start point (tsp) 73 nt upstream from the translational initiator. Both in vitro transcription assays and transcription of a chimeric gene in intact hepatoma cells indicated that the sequence located immediately upstream from the tsp contained a promoter. Several putative cis-regulatory elements, including a TATA box and a C/EBP-binding site were found within the 250 bp preceding the tsp.
从大鼠基因组文库中分离出编码胰岛素受体酪氨酸激酶抑制剂(PP63)的基因(PP63)。通过Southern印迹分析和序列数据(即外显子+边界)推断内含子/外显子结构。定位于11号染色体的PP63基因跨度约为8 kb,包含7个外显子,由6个大小不同的内含子分隔。所有边界均与供体和受体剪接位点的共有GT/AG序列匹配。引物延伸和S1作图实验用于确定转录起始点(tsp)位于翻译起始位点上游73 nt处。体外转录分析和完整肝癌细胞中嵌合基因的转录均表明,位于tsp紧上游的序列含有一个启动子。在tsp之前的250 bp内发现了几个推定的顺式调控元件,包括一个TATA盒和一个C/EBP结合位点。