ISOLATION OF TUMOR SUPPRESSOR GENES BY SUBRACTION LIBRARIES

控制末端分化的基因是一类抑制基因。 为了找出与植物末端分化相关的基因 两个CDNA文库正在构建中。第一 文库来自R.1细胞，这些细胞对血清没有反应而分化 或转化生长因子(TGF)-β。第二个来自S.6细胞 用转化生长因子-β诱导分化。去调查 这些细胞中基因表达的差异，一种消减杂交 我们会采取行动。简而言之，体外合成的RNA来自一个文库 而来自另一个文库的cdna将被杂交以减去共同的 消息，并且唯一的单链消息将被亚克隆以创建 第三，“减法”图书馆。来自差减文库的克隆可以 然后测试它们诱导分化的能力和/或 抑制正常人或肿瘤性人支气管上皮的致瘤性 细胞系。

Genes controlling terminal differentiation are one class suppressor genes. In order to identify genes associated with the terminal differentiation of epithelial cells, two CDNA libraries are being constructed. The first library is from R.1 cells which do not differentiate in response to serum or transforming growth factor (TGF)-beta. The second is from S.6 cells treated with TGF-beta, which induces their differentiation. To investigate differences in gene expression in these cells, a subtractive hybridization approach will be taken. Briefly, in vitro synthesized RNA from one library and cDNA from the other library will be hybridized to subtract out common messages, and unique single-stranded messages will be subcloned to make a third, "subtracted" library. The clones from the subtracted library can then be tested for their ability to induce differentiation and/or to suppress tumorigenicity in normal or neoplastic human bronchial epithelial cell lines.