BIOCHEMICAL GENETICS OF MALE SEXUAL DETERMINATION

我们计划继续研究哺乳动物的性别决定和 利用小鼠进行实验观察的辨证分型 性别模糊患者的临床相关性研究。第一个焦点 最近将出现性别决定的Y染色体候选基因 由大卫·佩奇博士(Zfy)克隆。这个克隆的DNA序列预测了一种 锌指，DNA结合蛋白，具有可能的调节特性。我们 将通过表达它来测试它在性别决定中的作用，以及它的 反义，在转基因小鼠中寻找性别逆转。我们有 开发了一种我们认为是一种新的克隆策略来克隆 DNA将对来自该蛋白质的积极调控信号做出反应 酵母。这种方法可能使我们能够识别常染色体或X连锁基因 参与性别决定。我们还将在早期研究Zfy的表达 胚胎和在分化的小鼠性腺组织中，将寻找 小鼠两种Y染色体的结构变异 在C56BL/6J背景下的功能有所不同。我们努力侦测 小鼠早期胚胎的转录本将使用聚合酶链式反应 它有可能检测到微量的转录。我们有 已经证明Y染色体克隆在 阐明患者的性别歧义问题。此外，分析 Y染色体的重组，比如发生在XX男性身上的 帮助我们阐明减数分裂过程中的染色体机制。因此，我们 相信我们对性别决定的研究都增加了我们的 了解哺乳动物的发育并提供临床上有用的信息 在遗传学和内分泌诊所中看到的帮助患者的材料。

We plan on continuing our studies of mammalian sex determination and differentiation using mice for experimental observations followed by clinical correlations in patients with sexual ambiguity. The first focus will be on the Y chromosomal gene candidate for sex determination recently cloned by Dr. David Page (Zfy). The DNA sequence of this clone predicts a zinc finger, DNA binding protein with probable regulatory properties. We will test its role in sex determination by expression of it, and its antisense, in transgenic mice in order to look for sex reversal. We have developed what we believe is a novel cloning strategy to clone stretches of DNA that will respond to positive regulatory signals from this protein in yeast. This approach may allow us to identify autosomal or X-linked genes involved in sex determination. We will also study Zfy expression in early embryos and in differentiating gonadal tissues of mice and will search for variation in its structure between tow kinds of mouse Y chromosomes which function differently on the C56BL/6J background. Our efforts to detect transcripts in early mouse embryos will use the polymerase chain reaction which has the potential to detect minute amounts of transcription. We have already demonstrated that Y chromosomal clones are very useful in elucidating problems of sexual ambiguity in patients. In addition, analysi of reorganizations of the Y chromosomes such as those that occur in XX male help us to elucidate chromosomal mechanisms during meiosis. Thus, we believe that our studies of sex determination both increase our understanding of mammalian development and provide clinically useful material to aid patients seen in genetics and endocrine clinics.