Cloning and characterization of a novel upstream untranslated exon of the mouse Fgf-1 gene.
Cloning and characterization of a novel upstream untranslated exon of the mouse Fgf-1 gene.
复制标题
小鼠 Fgf-1 基因的新型上游非翻译外显子的克隆和表征。
DOI：
10.1016/s0378-1119(96)00433-7
复制
发表时间：
1996
期刊：
影响因子：
3.5
通讯作者：
Chiu,IM
中科院分区：
文献类型：
--
作者：
Hackshaw,KV;Furlow,ZM;Chiu,IM
成纤维细胞生长因子1 （Fibroblast growth factor 1， FGF-1或aFGF）是肝素结合生长因子的原型成员，在体内具有血管生成的能力。FGF-1与动脉粥样硬化、癌症、伤口修复和炎症性自身免疫性疾病有关。作为了解FGF-1在炎症和癌症发病机制中的作用的一部分，我们已经采取了分离和表征小鼠FGF-1基因的步骤。Southern blotting和序列分析显示，Fgf-1在人类、小鼠、仓鼠、大鼠和牛的编码区和上游非翻译区具有相当大的保守性。利用从大鼠前列腺特异性Fgf-1 cDNA的5 ' -未翻译外显子衍生的引物，通过PCR从小鼠基因组DNA中扩增出一个220 bp的产物。序列分析表明，该扩增子与大鼠FGF-cDNA序列对应区域有80%的相似性。从该扩增子和Fgf-1编码区设计的引物用于分离跨越整个小鼠Fgf-1基因的多个重叠基因组克隆。对这一新颖的5'-未翻译外显子上游的基因组序列进行测序分析，未发现典型的TATA， CCAAT序列。似乎FGF-1的多个非翻译外显子的出现是该基因在物种间高度保守的主题。
Fibroblast growth factor 1 (FGF-1 or aFGF), is the prototype member of the heparin-binding growth factors which are capable of angiogenesis in vivo. FGF-1 has been implicated in atherosclerosis, cancer, wound repair and inflammatory autoimmune diseases. As part of an effort to understand the role of FGF-1 in the etiopathogenesis of inflammation and cancer, we have undertaken steps to isolate and characterize the mouse Fgf-1 gene. Southern blotting and sequence analysis displayed considerable conservation within the coding and upstream untranslated regions of Fgf-1 in human, mouse, hamster, rat and bovine. By using primers derived from the 5′-untranslated exon of a rat prostate-specific Fgf-1 cDNA, a 220-bp product was amplified from mouse genomic DNA via PCR. Sequence analysis of this amplicon showed that there was 80% similarity with the corresponding region of the rat FGF-cDNA sequence. Primers designed from this amplicon and the Fgf-1 coding region were used to isolate multiple overlapping genomic clones spanning the entire mouse Fgf-1 gene. Sequencing analysis of the genomic sequence upstream from this novel 5'-untranslated exon did not reveal typical TATA, CCAAT sequences. It appears that the occurrence of multiple untranslated exons for FGF-1 is a highly conserved theme for this gene across species.