GENETIC BASIS OF MAMMALIAN SEX DETERMINATION

这项研究项目的目标是开发一张物理地图 小鼠Y染色体。具体地说，通过我们建议的研究，我们将： 1)创建200个分布的序列标记位点(STSS)的集合 在小鼠Y染色体的常染色质部分，2)使用 STSS来表征一组缺失-听力Y染色体，以及 3)使用STSS创建有序的酵母和/或细菌阵列 跨越常着丝点的人工染色体(YAC或BAC)克隆 Y染色体的一部分。 双变量荧光激活法分离Y-特异性物质 男性和女性的染色体分选和/或消减杂交 基因组DNA，将用于创建小插入的STS源库 大小(500个碱基对)。三四百个特定于Y的克隆将是 测序，目标是产生至少200个单拷贝，Y- 特定的STS引物对。然后，STSS胜利被用来描述一个 一系列带有缺失的Y染色体，决定了两个STSS的顺序 和删除断点，从而创建 Y。STSS还将用于分离和排序连续的系列 跨越Y染色体的YAC或BAC克隆。总而言之，建议的 实验将使我们更深入地了解这种结构和 Y染色体的进化。此外，通过提供直接的分子 Y上基因克隆的切入点，建议进行实验 将为性别决定的研究提供新的见解，男性 生育能力和疾病(如高血压和性母细胞瘤) 显示出与某些Y连锁等位基因的关联。

The goal of this research project is to develop a physical map of the mouse Y chromosome. Specifically, through our proposed research we will: 1) create a collection of 200 sequence tagged sites (STSs) distributed throughout the euchromatic portion of the mouse Y chromosome, 2) use the STSs to characterize a collection of deletion-hearing Y chromosomes, and 3) use the STSs to create an ordered array of yeast and/or bacterial artificial chromosome (YAC or BAC) clones spanning the euchromatic portion of the Y chromosome. Y-specific material isolated by bivariate fluorescence activated chromosome sorting, and/or subtractive hybridization of male and female genomic DNA, will be used to create an STS source library of small insert size (500 base pairs). Three to four hundred Y-specific clones will be sequenced with the goal of generating at least 200 single copy, Y- specific, STS primer pairs. The STSs win then be used to characterize a series of deletion-bearing Y chromosomes, giving order to both the STSs and the deletion breakpoints, and thereby creating a physical map of the Y. The STSs will also be used to isolate and order a contiguous series of YAC or BAC clones spanning the Y chromosome. Together, the proposed experiments will give us a deeper understanding of the structure and evolution of the Y chromosome. Moreover, by providing a direct molecular entry point for cloning genes borne on the Y, the proposed experiments will afford new insights into the study of sex determination, male fertility, and illnesses (such as hypertension and gonadoblastoma) that show an association with certain Y-linked alleles.