PED GENE ACTION IN DEVELOPMENT AND REPRODUCTION

描述：(从申请者的描述中扫描)： 植入前的哺乳动物胚胎，以及由 这些胚胎是由植入前表达的基因调控的 发展时期。一个例子是小鼠着床前胚胎 发育(Ped)基因。Ped基因具有多效性作用。 胚胎卵裂分裂、足月存活率、出生体重和断奶 重量。Ped基因产物已被确定为主要的 组织相容性复合体(MHC)编码Lb类蛋白Qa-2。小鼠胚胎 表达Qa-2蛋白的人切割速度更快，有更高的机会 存活到出生，并产生出生体重和断奶体重较高的幼崽 而不是不表达Qa-2的胚胎。令人费解的是，一个 胚胎细胞表面的蛋白质可以发出信号，让胚胎在 比缺少这种蛋白质的胚胎速度更快。这一发现表明 PED基因产物是一种MHC类Ib蛋白，提供了什么结构的线索 为了解开这一谜团而进行检查的特征。首先，非聚合体的多肽 结合Qa-2蛋白将通过膜技术进行鉴定 预富集-毛细管电泳串联质谱 (MPC-CE-MSfMS)。第二，与Qa-2结合的辅助蛋白的类型 有助于调节其功能的蛋白质将通过使用酵母进行鉴定 双杂交系统。Ped基因的重要多效性作用 繁殖的成功使人们希望搜索与此相关的人类同源物 吉恩。人类MHC(人类白细胞抗原)DNA全序列的公布 复合体)，以及最近关于编码在 人类白细胞抗原复合体，表明最符合逻辑的假设是人类同源物 Qa-2为人类白细胞抗原-G。这一假设将在第三个具体目标中得到检验 使用人类白细胞抗原G转基因小鼠系统。

DESCRIPTION: (Scanned from the applicant's description): The survival of preimplantation mammalian embryos, and the health of offspring arising from these embryos, is regulated by genes expressed during the preimplantation period of development. One example is the mouse Preimplantation embryo development (Ped) gene. The Ped gene has pleiotropic effects on the rate of embryonic cleavage division, survival to term, birth weight, and weaning weight. The Ped gene product has been identified as the major histocompatibility complex (MHC) encoded class lb protein Qa-2. Mouse embryos that express Qa-2 protein cleave at a faster rate, have a higher chance of survival to birth, and give rise to pups with higher birth and weaning weights than embryos that do not express Qa-2. The mystery is how the presence of a protein on the embryonic cell surface can signal the embryo to cleave at a faster rate than an embryo that is missing this protein. The finding that the Ped gene product is a MHC class Ib protein provides clues about what structural features to examine to solve this mystery. First, the nonameric peptides that bind to Qa-2 protein will be identified by using the technique of membrane preconcentration-capillary electrophoresis-tandem mass spectrometry (mPC-CE-MSfMS). Second, the types of accessory proteins that bind to Qa-2 protein to help mediate its function will be identified by using the yeast two-hybrid system. The important pleiotropic effects of the Ped gene on reproductive success make it desirable to search for a human homolog of this gene. The publication of the complete DNA sequence of the human MHC (HLA complex), along with recent literature on MHC class lb proteins encoded in the HLA complex, suggest that the most logical hypothesis is that a human homolog of Qa-2 is HLA-G. This hypothesis will be tested in the third specific aim by using an HLA-G transgenic mouse system.