BIOCHEMICAL GENETICS OF MALE SEXUAL DETERMINATION

我们计划继续研究哺乳动物的性别决定， 分化使用小鼠进行实验观察， 性模糊患者的临床相关性。 第一焦点 将在Y染色体基因的候选人性别决定最近 由大卫佩奇博士（Zfy）克隆。 这个克隆的DNA序列预测了一个 锌指，可能具有调节特性的DNA结合蛋白。 我们 将测试其在性别决定中的作用， 反义，在转基因小鼠，以寻找性别逆转。 我们有 开发了我们认为是一种新的克隆策略， DNA将对来自这种蛋白质的阳性调节信号做出反应， 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 与性别决定有关 我们还将研究Zfy在早期乳腺癌中的表达， 胚胎和分化性腺组织的小鼠，并将寻找 两种小鼠Y染色体之间的结构差异， 在C56 BL/6 J背景上的功能不同。 我们的努力， 早期小鼠胚胎中的转录本将使用聚合酶链反应， 其具有检测微量转录的潜力。 我们有 已经证明Y染色体克隆在 阐明了患者的性别模糊问题。 此外，分析 Y染色体的重组，例如发生在XX男性中的重组， 帮助我们阐明减数分裂中的染色体机制。 因此我们 相信我们对性别决定的研究， 了解哺乳动物的发育，并提供临床上有用的 材料，以帮助病人看到在遗传学和内分泌诊所。

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.