HUMAN GENOME CLONING AND ISOLATION OF SPECIFIC DNAS IN YEAST

工作总结：染色体稳定小组在焦油克隆方面的实质性进展为建立一个单独的、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焦油克隆方法。我们与西南得克萨斯大学的罗恩·沃尔特开展了一项合作，从剑鱼中开发酵母人工文库(YAC)，因为这些鱼是DNA修复和肿瘤抑制控制的优秀全动物遗传模型。我们还开始分离已知肿瘤抑制基因已被定位的特定基因周围的区域(Xmrk和CDKn2区域)。我们正在改进cDNA的焦油克隆，以使用我们的酵母功能分析来检查各种癌症组织中表达的基因。在其他研究中，我们已经表明，从人类细胞系和癌症组织中焦油克隆P53基因是一个准确的过程，基于酵母的系统可以用于表征突变的P53基因。根据人P53的焦油克隆结果，我们启动了一项从剑鱼中克隆和检测P53的计划，然后在我们的酵母系统中检测不同诱导物中的P53‘S。

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.