IMMUNOGENETICS OF SPERMATOZOA, EGGS, AND EMBRYOS

小鼠17号染色体上的T/t复合体，包括主要的 组织相容性复合体(MHC)，还含有多个突变基因 会影响精子的分化，以及其他功能 在早期胚胎发育过程中。因此，它一直在密集地 作为模型系统被研究用于分析免疫生物学和 分化以及它们的基因控制。然而，直到最近， Tau复合体的分子生物学仅限于染色体行走， 标记识别和随机筛选。这一阶段现在已经 完成了。来自这个实验室的新结果，是通过一个 基因和分子技术的结合表明，MHC 成为该网站对tau-Complex的主要兴趣，并提供了工具 研究哺乳动物体内调节发育的基因复合体。五 在新克隆的6个男性生殖细胞表达基因中，MHC 显示野生型与tau突变型睾丸的表达变化。就.而言 早期胚胎发育，四个隐性的、全致死的早期胚胎 在散布着MHC基因的基因中，有两个基因被定位得更精确： TW5在基因上与H-2K密不可分，也是最早的演技之一 哺乳动物致死基因T12位于Tl区或未克隆的 TL和QAlpha区域之间的“间隙”。表达的早期胚胎 转录因子Oct-4也被定位到TL和 QAlpha，初步研究表明它很可能是T12。 这个项目提出了三个相辅相成的战略来定义 这些有趣的tau复合体基因的分子和生物学功能。 (1)从转录水平对克隆的睾丸表达基因进行分析， 翻译和蛋白质水平。最终，我们将使用转基因技术 技术来研究它们的主导功能。(2)克隆鉴定和作图 基因，来自我们的实验室和其他实验室，转录自 在tau复合体中，并在小鼠早期胚胎中表达。我们会 把注意力集中在那些有tau致死突变的候选基因上。 目前，这包括Oct-4和其他六个早期胚胎tau/tau 复杂的编码基因。(3)重组遗传分析 Tau/tau复合区支持分子和功能分析。 我们的长期目标是了解分子机制和 细胞承诺的基因控制。

The T/t complex on mouse chromosome 17 which includes the major histocompatibility complex (MHC), also contains a number of mutant gene that affect the differentiation of spermatozoa, and others that function during early embryonic development. Thus, it has been intensively investigated as model system for analyzing both immunobiology and differentiation, and their genetic control. Until recently, however, the molecular biology of the tau-complex was limited to chromosomal walking, marker identification, and random screening. This phase has now been accomplished. New results from this laboratory, obtained with a combination of genetic and molecular techniques have shown that the MHC has become the site major interest in the tau-complex and provided the tools to investigate a gene complex that regulates development in a mammal. Five out of six newly cloned male germ-cell-expressed genes residing in the MHC show expression changes in wild type versus tau-mutant testis. In terms of early embryo development, of the four recessive, early embryo tau-lethal genes interspersed with MHC genes , two have been more precisely located: tw5 is genetically inseparable from H-2K, and one of the earliest acting mammalian lethals, t12, is located either in the TL region or the uncloned "gap" between the TL and Qalpha regions. The early-embryo expressed transcription factor, Oct-4, has also been mapped to the gap between TL and Qalpha, and preliminary studies indicate it may well be t12. This project proposes three complementary strategies to define the molecular and biological function of these interesting tau-complex genes. (1) Analyze the cloned testis-expressed genes at the transcriptional, translational, and protein level. Ultimately, we will employ transgenic technology to study their dominant function. (2) Identify and map cloned genes, both from our laboratory and others, that are transcribed from within the tau-complex, and expressed in early mouse embryos. We will concentrate on those that are candidates for the tau-lethal mutation. Currently, this includes Oct-4 and six other early embryonic tau/tau complex encoded cDNAs. (3) Recombinational genetic analysis of the tau/tau complex region to support the molecular and functional analysis. Our long- term objective is to understand the molecular mechanisms and genetic control of cellular commitment.