DEVELOPMENT OF EXPRESSION CLONING SYSTEM FOR ONCOGENE CDNAS

设计了一种表达克隆策略来分离角质形成细胞 生长因子(KGF)受体基因的自分泌 变换循环。NIH/3T3成纤维细胞，分泌这种上皮细胞 细胞特异性有丝分裂原，将角质形成细胞表达 C DNA文库。在确认的几个转化灶中，有一个 证明获得了特异的高亲和力KGF结合位点。 相关成纤维细胞生长因子的结合竞争模式 (FGFs)表明该受体与酸性成纤维细胞生长因子有较高的亲和力 (a成纤维细胞生长因子)和KGF。回收的4/2-kb的cdna编码一个 预测的膜扫描酪氨酸激酶与之相关但不同 碱性成纤维细胞生长因子(BFGF)受体。这种表达克隆方法可能是 一般适用于构成限制基因的分离 有丝分裂信号通路中的步骤。

An expression cloning strategy was devised to isolate the keratinocyte growth factor (KGF) receptor cDNA by creation of an autocrine transforming loop. NIH/3T3 fibroblasts, which secrete this epithelial cell-specific mitogen, were transfected with a keratinocyte expression cDNA library. Among several transformed foci identified, one demonstrated the acquisition of specific high affinity KGF binding sites. The pattern of binding competition by related fibroblast growth factors (FGFs) indicated that this receptor had high affinity for acidic FGF (aFGF) as well as KGF. The rescued 4/2-kb cDNA was shown to encode a predicted membranespanning tyrosine kinase related to but distinct from the basic FGF (bFGF) receptor. This expression cloning approach may be generally applicable to the isolation of genes that constitute limiting steps in mitogenic signalling pathways.