BIOCHEMICAL GENETICS OF MALE SEXUAL DETERMINATION

我们试图了解与细胞分化有关的分子事件 哺乳动物的雄性胚胎性腺。这样的知识应该是有益的。 改进人类双性人的诊断，可能还有产前诊断 以及增加我们对正常人类的理解 发展。我们的研究将同时使用老鼠模型和人类材料。 对小鼠的研究将集中在近臂的遗传基因座上。 与小鼠Y染色体相互作用的17号染色体(它也有 变异体)来尝试并阐明这些常染色体基因座如何与Y 染色体和其他常染色体基因座对正常睾丸的影响 发展。在17号染色体上的基因座中，可能有一些包含 克雷特小卫星(BKM)的性别特异序列。因为确定了 17号染色体这一区域的大量缺失(T-OR1和T-HP)可导致 两性畸形，我们将确定这些缺失是否会改变 原位检测17号染色体上BKM-序列的数量 杂交。我们将使用BKM探针来寻找包含BKM的mRNAs 在小鼠和人类胎儿性腺和适当的其他组织中的序列。 小鼠17号染色体近端和lambda的Cosmid克隆 来自人类X染色体的克隆将用BKM探针进行筛选。 阳性克隆的亚群将用于筛查胎儿性腺RNA。 已获得的人类Y染色体克隆和新获得的Y染色体克隆 通过PERT和染色体“HOP”技术，将被用于研究XX雄性和 他们的父母试图确定Y染色体的来源 通常在这类XX男性身上发现DNA。人类材料方面的工作也将 利用人类Y染色体克隆在男性中寻找表达的Y序列 胎儿性腺。与小鼠或人类男性胎儿相对应的基因组克隆 筛选cDNA性腺特异性表达序列 用鉴定的探针构建文库，并用该克隆进行筛选 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).