MOLECULAR BASIS OF THE PED GENE PHENOTYPE

PED 基因表型的分子基础

• 批准号: 2673744
• 负责人: CAROL M WARNER
• 金额: $ 25.67万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-05-01 至 2000-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2673744
• 关键词: MHC class I antigen; developmental genetics; early embryonic stage; embryo implantation; embryogenic cleavage; enzyme linked immunosorbent assay; flow cytometry; genetically modified animals; glycosylphosphatidylinositols; in situ hybridization; laboratory mouse; linkage mapping; mammalian embryology; microinjections; molecular cloning; monoclonal antibody; northern blottings; nucleic acid sequence; phenotype; pleiotropism; polymerase chain reaction; regulatory gene; southern blotting; subtraction hybridization; tissue /cell culture; weaning

The preimplantation period of mammalian development is genetically controlled by both maternal and embryonic genes. One embryonic gene, Ped (Preimplantation embryo development), determines the rate (fast or slow) at which preimplantation mouse embryos cleave. In addition, the Ped gene has well-defined model system for the system for the study of early mammalian embryonic development and reproductive success. The molecular mechanisms by which the Ped gene product, a class I major histocompatibility complex (MHC) protein, the Qa-2 antigen, confers the Ped gene phenotype, will be evaluated. Only embryos expression Qa-2 antigen cleave at a fast rate. Four similar genes, Q6, Q7, Q8, and Q9 encode the Qa-2 antigen. The relative contribution of each of these genes to the various manifestations of Ped gene phenotype is unknown, but it has been shown, by the analysis of Q9 transgenic mice, that the Q9 gene controls the rate of cleavage division of early embryos. The contribution of the Q9 gene to the other aspects of Ped gene phenotype, litter size, birth weight, and weaning weight, will be analyzed. In addition, new lines of Q9 transgenic mice will be evaluated to determine whether chromosomal location or copy number of the Q9 gene affect Ped gene phenotype. Next, the type of membrane linkage of the Qa-2 antigen to the embryonic cell surface will be studied, by using Q9 exon-spliced gene constructs, to determine whether a glycosylphosphatidylinositol (GPI) linkage is required for Qa-2 antigen to signal embryos to cleave at a fast rate. The possible contribution to Ped gene phenotype of the other Qa-2 encoding genes, Q6, Q7, and Q8, will also be evaluated. Subtractive hybridization experiments will be used to classify any as yet unidentified genes that may contribute to the Ped gene phenotype. Also to be analyzed are the distribution of Qa-2 antigen on the embryonic cell surface and whether cross-linking of Qa-2 antigen on the embryonic cell surface and whether cross-linking of Qa-2 antigen on the embryonic cell surface can activate embryos to cleave at a faster rate. Finally, studies will be conducted to ascertain to their Qa-2 negative littermates, occurs. The finding that Ped gene product is a class I MHC protein is of fundamental interest because this suggests that the MHC class I family of proteins may be central importance to cell-cell interactions in development and reproduction as well as to cell-cell interactions in the immune response.

哺乳动物发育的植入前阶段是遗传的 由母体基因和胚胎基因共同控制。一个胚胎基因，Ped (植入前胚胎发育)，决定了速度(快或慢) 植入前的小鼠胚胎在这个位置分裂。此外，Ped基因 具有良好定义的模型体系，便于早期的系统研究 哺乳动物胚胎发育和繁殖成功。分子 Ped基因产物的机制，I类专业 组织相容性复合体(MHC)蛋白，Qa-2抗原，赋予 PED基因表型，将进行评估。只有胚胎表达Qa-2 抗原以很快的速度分裂。四个相似的基因，Q6、Q7、Q8和Q9 编码Qa-2抗原。其中每一项的相对贡献 基因对Ped基因的各种表现表型尚不清楚，但 对Q9转基因小鼠的分析表明，Q9 基因控制早期胚胎的卵裂分裂速度。这个 Q9基因对Ped基因表型其他方面的贡献 将分析产仔数、初生重和断奶重。在……里面 此外，将对Q9转基因小鼠的新品系进行评估，以确定 Q9基因的染色体位置或拷贝数是否影响Ped 基因表型。接下来，Qa-2抗原的膜连接类型 利用Q9外显子剪接对胚胎细胞表面的影响 基因构建，以确定糖基磷脂酰肌醇是否 Qa-2抗原需要(GPI)连锁才能发出胚胎分裂的信号 以很快的速度。中国人Ped基因表型的可能贡献 其他Qa-2编码基因Q6、Q7和Q8也将被评估。 消减杂交实验将被用来对尚未发现的 可能与Ped基因表型有关的未知基因。也是 有待分析的是qa-2抗原在胚胎细胞上的分布。 QA-2抗原在胚胎细胞表面的交联性 QA-2抗原在胚胎细胞表面的交联性 表面可以激活胚胎以更快的速度分裂。最后， 将进行研究，以确定他们的QA-2阴性 产仔，发生。发现Ped基因产物是一种I类MHC 蛋白质是最重要的，因为这表明MHC I类蛋白家族可能对细胞-细胞起核心作用 发育和繁殖中的相互作用以及细胞与细胞之间的相互作用 免疫反应中的相互作用。