BIOCHEMICAL GENETICS OF MALE SEXUAL DETERMINATION

我们试图了解参与分化的分子事件， 哺乳动物雄性胚胎性腺。 这样的知识应该是有益的 用于改进人类中间性的诊断， 以及增加我们对正常人的了解 发展 我们的研究将使用小鼠模型和人类材料。 在小鼠中的研究将集中在近臂的遗传位点， 与小鼠Y染色体相互作用的17号染色体（也具有 试图阐明这些常染色体基因座如何与Y染色体相互作用， 染色体和其他常染色体基因座在正常睾丸发育中的作用 发展 在17号染色体上的基因座中， 带状金环蛇小卫星（Bkm）性别特异性序列。 因为某些 17号染色体这一区域的大缺失（T-Or 1和T-Hp）可导致 我们将确定这些缺失是否改变了 通过原位杂交在17号染色体上检测到的BKm序列的量 杂交方法 我们将使用Bkm探针寻找含有Bkm的mRNA 小鼠和人胎儿性腺和适当的其它组织中的序列。 小鼠17号染色体近端区域的粘粒克隆和λ 用Bkm探针筛选来自人X染色体的克隆。 阳性克隆的亚部分将用于筛选胎儿性腺RNA。 先前获得的人类Y染色体克隆和获得的新克隆 通过PERT和染色体“跳跃”技术，将用于研究XX男性和 他们的父母试图确定Y染色体的来源 DNA通常发现在这样的XX男性。 对人体材料的研究也将 使用人类Y染色体克隆寻找男性中表达的Y序列 胎儿性腺 对应于小鼠或人类雄性胎儿的基因组克隆 通过筛选cDNA寻找性腺特异表达序列 文库，并使用cDNA克隆进行筛选 基因组克隆（当探针不是唯一序列时）。

We seek to understand molecular events involved in the differentiation of the mammalian male embryonic gonad. Such knowledge should be of benefit for improved diagnosis, and possibly prenatal diagnosis, of human intersex conditions as well as increasing our understanding of normal human development. Our studies will use both mouse models and human material. Studies in mice will focus on genetic loci in the proximal arm of chromosome 17 which interact with the mouse Y chromosome (which also has variants) to try and elucidate how these autosomal loci interact with the Y chromosome and other autosomal loci in contributing to normal testicular development. Among the loci on chromosome 17 may be those containing Banded krait minor satellite (Bkm) sex specific sequences. Since certain large deletions (T-Or1 and T-Hp) in this area of chromosome 17 can cause hermaphroditism, we will determine whether or not these deletions alter the amounts of Bkm-sequences detected on chromosome 17 by in situ hybridization. We will use the Bkm probe to look for mRNAs containing Bkm sequences in mouse and human fetal gonads and appropriate other tissues. Cosmid clones from the proximal region of mouse chromosome 17 and lambda clones from the human X chromosome will be screened with a Bkm probe. Subportions of positive clones will be used to screen fetal gonad RNA. The human Y chromosomal clones previously obtained, and new ones obtained by PERT and chromosome "hop" techniques, will be used to study XX males and their parents in an attempt to determine the source of the Y chromosomal DNA usually found in such XX males. The work on human material will also use human Y chromosome clones to look for expressed Y sequences in the male fetal gonad. Genomic clones corresponding to mouse or human male fetal gonad specific expressed sequences will be sought by screening cDNA libraries with the identified probe and using the cDNA clone to screen genomic clones (when the probe is not unique sequence).