YEAST BTNLP AND HUMAN CLN3P IN YEAST

一种可能与巴顿S病有关的人类基因最近 都被克隆了。这是一种非常严重的疾病，会导致神经退化。 以及许多儿童的死亡。该缺陷的性质导致了 疾病尚不为人所知。这种人类基因，CLN3，在 酵母，BTN1。BTN1零突变体是可行的，但确实有微妙的 表型：在某些条件下，突变体有些 对D-(-)three-2-amino-1-[p-nitropheny1]-1，3-propanediol的抗性 (ANP)。调查人员建议对巴顿S病进行调查 酵母和BTN1基因作为模型系统。私家侦探S将调查 正常和突变形式的Cln3p和 Btnip，并将确定蛋白质的关键区域。这个 将使用GFP或表位确定Btnip的亚细胞位置 标签。利用双杂交技术将发现与Btnip相互作用的蛋白质 系统，以及通过使用GST-Ttnip分离可能的复合体 多组氨酸标记的Fusion或Btnip。将进行基因筛查 为了寻找其他对APN具有抗性的突变体，寻找btn1的抑制子， 并找到对BTN1具有合成致命性的突变。

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.