US-France Cooperative Research: Comparison of Two Double Strand Break Mediated Recombination Events

该奖项将支持真核生物领域的合作研究。 布兰代斯大学的詹姆斯·哈伯博士和 Bernard Dujon，巴斯德研究所，巴黎，法国。 的目标 本课题是对两种不同重组方式进行比较和对比 酵母系统 双链断裂（DSB）在启动重组中的作用 已经在实验中得到了很好的证实， 诱导重组到“靶向”转化， 限制性内切酶切割的DNA。 本工程 研究人员将比较HO-和W-内切酶 在酵母中启动DSB重组，使用遗传和物理 分析方法. 这项工作将使用半乳糖进行- 或磷酸调节基因以控制核酸酶的表达。 将主要使用含有以下的质粒测定纯化： 大肠杆菌LACZ基因的转录和非转录拷贝， 其中转录的拷贝被核酸内切酶破坏 识别序列 两位研究人员都进行了重大的 重组系统的工作，拟议的项目将 从他们的专业知识中受益匪浅。 这种合作 为研究重组事件提供了一种独特的方法， 发生在真核细胞的不同细胞器中。

This award will support collaborative research in eukaryotic genetics between Dr. James Haber, Brandeis University, and Dr. Bernard Dujon, Pasteur Institute, Paris, France. The objective of the project is to compare and contrast two different recombination systems of yeast. The role of double-strand breaks (DSB) in initiating recombination has been well established in experiments ranging from X-ray induction of recombination to the "targeted" transformation of restriction endonuclease-cleaved DNA. In this project, the researchers will compare and contrast HO- and w-endonuclease initiated DSB recombination in yeast, using genetic and physical methods of analysis. The work will be carried out using galactose- or phosphate-regulated genes to control expression of the nuclease. Recombination will be primarily assayed using a plasmid containing transcribed and untranscribed copies of the E.coli LACZ gene, where the transcribed copy is disrupted by the endonuclease recognition sequence. Both researchers have carried out significant work on recombination systems, and the proposed project will benefit greatly from their combined expertise. This collaboration offers a unique approach to the study of recombination events that occur in different organelles of the eukaryotic cell.