YEAST BTNLP AND HUMAN CLN3P IN YEAST

最近，一种可能导致巴顿病的人类基因， 被克隆了 这是一种非常严重的疾病，会导致神经退化 许多儿童死亡。 造成缺陷的性质 疾病未知。这种人类基因CLN 3在人类中有一个同源基因， 酵母，BTN 1. 一个btn 1无效突变体是可行的，但有一个微妙的 在某些条件下，突变体在某种程度上 抗D-（-）三-2-氨基-1-[p-硝基苯]-1，3-丙二醇 （ANP）。 研究人员建议使用 酵母和BTN 1基因作为模型系统。私家侦探s将进行调查 正常和突变改变形式的Cln 3 p的表达， Btnip，并将确定蛋白质的关键区域。的 Btnip的亚细胞定位将使用GFP或表位 标签. 与Btnip相互作用的蛋白质将使用双杂交技术发现。 系统，以及通过使用GST-Ttnip分离可能的复合物 融合物或用聚组氨酸标记的Btnip。基因筛查将在 寻找其他抗APN的突变体，寻找btn 1的抑制因子， 并找到与BTN 1合成致死的突变。

A human gene probably responsible for Batten s disease has recently been cloned. This is a very serious disease causing neurodegeneration and death in many children. The nature of the defect causing the disease is not known. This human gene, CLN3, has a homologue in yeast, BTN1. A btn1 null mutant is viable, but does have a subtle phenotype in that under some conditions the mutant is somewhat resistant to D-(-)three-2-amino-1-[p-nitropheny1]-1,3-propanediol (ANP). The investigators propose to investigate Batten s disease using yeast and the BTN1 gene as a model system. The P.I. s will investigate expression of normal and mutationally altered forms of Cln3p and Btnip, and will determine critical regions of the proteins. The subcellular location of Btnip will be determined using GFP or epitope tags. Proteins interacting with Btnip will be found using the two-hybrid system, as well as by isolating possible complexes using a GST-Ttnip fusion or Btnip tagged with poly-histidine. Genetic screens will be done to find other mutants resistant to APN, to find suppressors of btn1, and to find mutations that are synthetically lethal with btn1.