HUMAN GENOME CLONING AND ISOLATION OF SPECIFIC DNAS IN YEAST

工作总结：染色体稳定性小组在TAR克隆方面取得的重大进展，为开发一个单独的、独立的基因分离单位提供了条件，该单位致力于改进哺乳动物组织的TAR克隆方法。 我们开始与西南德克萨斯大学的罗恩沃尔特合作，从剑尾鱼中开发酵母人工文库（YAC），因为这些鱼是DNA修复和肿瘤抑制基因控制的优秀全动物遗传模型。我们还开始分离特定基因周围的区域，其中已知的肿瘤抑制基因已被定位（Xmrk和Cdkn2区域）。利用我们的酵母功能分析，对cDNA的TAR克隆进行了改进，以检查来自各种癌症组织的表达基因。在其他研究中，我们已经表明，TAR克隆人细胞系和癌组织的p53 cDNA是一个准确的过程，酵母为基础的系统可以用来表征突变p53。 基于TAR克隆人p53的结果，我们已经启动了一个项目，从剑突中克隆和测试p53，然后在我们的酵母系统中测试来自不同诱导肿瘤的p53。

Summary of Work: The substantial advancements in TAR cloning in the Chromosome Stability Group provided for development of a separate, independent Gene Isolation Unit that devotes much of its efforts to refining TAR cloning approaches for mammalian tissues. We initiated a collaboration with Ron Walter, Southwest Texas University, to develop yeast artificial libraries (YAC) from the fish Xiphophorous, since these fish are excellent whole animal genetic models for DNA repair and tumor suppressor control. We have also started the isolation of regions surrounding specific genes where known tumor suppressor genes have been mapped (Xmrk and Cdkn2 regions). TAR cloning of cDNAs is being refined to examine expressed genes from various cancer tissues using our yeast functional assays. In other studies we have shown that TAR cloning of p53 cDNA from human cell lines and cancer tissue is an accurate process and that yeast-based systems can be used to characterize mutant p53s. Based on the results of the TAR cloning of the human p53, we have initiated a project to clone and test the p53 from Xiphophorous, and then to test p53's from different induced tumors in our yeast based system.