IMMUNOGENETICS OF SPERMATOZOA, EGGS, AND EMBRYOS

精子、卵子和胚胎的免疫遗传学

• 批准号: 3311359
• 负责人: DOROTHEA BENNETT
• 金额: $ 30.13万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 1976
• 资助国家: 美国
• 起止时间: 1976-05-01 至 1991-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3311359
• 关键词: HY antigen; antigens; cell differentiation; chromosome aberrations; chromosome complement; developmental genetics; early embryonic stage; egg /ovum; embryo /fetus; gene expression; genetic manipulation; genetic mapping; haploidy; homozygote; immunogenetics; laboratory mouse; mammalian embryology; membrane activity; molecular cloning; mutant; nucleic acid sequence; nucleic acid structure; sex chromosomes; sperm; spermatogenesis

The T/t complex on mouse chromosome 17 contains a number of mutant genes that function during early embryonic development, and others that affect the differentiation of spermatozoa. Thus it has been intensively investigated as a model system for analyzing differentiation and its genetic control. Until recently, however, the T/t complex was not approachable in terms of molecular biology since the genetic and molecular markers necessary for investigation at the level of DNA structure were not available. New results from this laboratory, obtained with a combination of genetic, serological, and molecular techniques have now provided the necessary material for intensive molecular analysis. We have defined 7 t-lethal genes as non-allelic but functionally related, and have accurate information on their location relative to one another and to other chromosome markers, including the TL, K, I, anbd D regions of the MHC. Four of the lethal genes are interspersed with MHC genes. We have generated an extensive panel of recombinant chromosomes, derived by crossing over between two different t-haplotypes, and defined their breakpoints. Fourteen of these are intra H-2 recombinants, with breakpoints defined by restriction fragment polymorphisms for 7 MHC probes. Thus for the first time the opportunity exists for the molecular investigation of a gene complex that regulates development in a mammal. This project proposes three complementary strageties to define the molecular organization of the mutant T/t complex and compare it to its wild type counterpart, and to identify and clone mutant genes in that region. (1) Chromosomal walking in a cosmid genomic library of homozygous t-w5 DNA, initiated with probes to H-2 class I genes and with K region probes that map very close (0-0.1 cM) to the t-w5 lethal mutation, will produce a restriction map of the MHC that can be compared to wold type. (2) A similar chromosomal walk in a cosmid genomic library of t-12/t-w5 DNA will permit a comparison of the restriction maps of the MHC of t-12 with t-w5. This comparison will be especially interesting because it will give information on evolutionary changes in DNA structure that can occur in the virtual absence of recombination. It also may help to pinpoint structural aberrations associated with the t-12 and t-w5 lethal mutations. (3) Cloning of chromosome 17 sequences expressed during spermatogenesis in normal and mutant testes to identify and map genes responsible for T/t complex phenotypes.

小鼠17染色体上的T/T复合物包含许多突变基因 在早期胚胎发展期间的功能，以及其他影响的 精子的分化。 因此，它一直是 作为分析分化及其ITS的模型系统调查 遗传控制。 但是，直到最近，T/T复合物还没有 自遗传和分子以来，就分子生物学而定 在DNA结构水平上进行调查所需的标记不是 可用的。 该实验室的新结果，结合了 遗传，血清学和分子技术现已提供 进行密集分子分析的必要材料。 我们定义了7 T致命基因作为非平行基因，但在功能上相关，并且具有准确的 有关其位置相对于彼此以及其他位置的信息 染色体标记，包括MHC的TL，K，I，ANBD D区域。 四个致命基因与MHC基因散布。 我们有 产生了大量的重组染色体，由 在两种不同的t-haplotypes之间越过，并定义了它们 断点。 其中十四个是H-2内重组剂，有 由7 MHC的限制片段多态性定义的断点 探针。 因此，第一次存在分子的机会 研究调节哺乳动物发育的基因复合物。 该项目提出了三个互补的散乱，以定义 突变体T/T复合物的分子组织并将其与野生进行比较 类型对应物，并在该区域识别和克隆突变基因。 （1）在纯合T-W5 DNA的宇宙基因组库中行走的染色体行走， 用探针对H-2 I类基因和K区域探针发起 地图非常接近（0-0.1 cm）到T-W5致命突变，将产生一个 可以将MHC与Wold类型进行比较的限制图。 （2）a 在T-12/T-W5 DNA的宇宙基因组库中类似的染色体步行将 允许将T-12的MHC的限制图与T-W5进行比较。 这种比较将特别有趣，因为它将给予 有关可能发生在DNA结构中进化变化的信息 虚拟缺乏重组。 它也可能有助于查明结构 与T-12和T-W5致命突变有关的畸变。 （3） 在精子发生过程中表达的17个染色体的克隆 正常和突变睾丸识别和映射负责T/T的基因 复杂的表型。