GENETIC STUDIES OF PREGNANCY-SPECIFIC B1 GLYCOPROTEIN

本建议的长期目标是了解功能 不同种类“妊娠特异性β1-糖蛋白”的作用 (SP1)“在人类妊娠中的作用”，以期加强对影响因素的认识 这影响到怀孕的确定和妇女的福祉 发育中的胎儿。这项提议的直接目标包括澄清 不同SP1cDNAs和基因的结构，表明 调节基因表达的因素，并定义潜在的 这些蛋白质的生理功能。最初，核苷酸 不同SP1cDNA的序列将被确定。基因专一性 将根据结构合成寡核苷酸探针 这些cDNA之间的差异。不同种类的人SP1蛋白 将通过体外表达的方式进行制备。特异性单抗 还将制备针对每一种SP1的(MAb)。影响经济增长的因素 RNA斑点印迹技术的表达。对妊娠的影响将是 使用胎盘组织和母体血清进行研究。组织特异性 SP1基因产物将在组织培养和使用cdna进行检测。 来自不同组织的文库。激素和其他物质的影响 分子将通过滋养层培养进行检测。SP1基因将是 关于内含子/外显子位置以及 使用以下技术的转录起始和终止位点 限制性内切酶图谱、Northern和Southern杂交分析，S1 核酸酶保护法、引物延伸法和核跑接法。 将通过染色体定位来研究SP1基因家族的组织 使用基因特异性探针和染色体行走。神经细胞的功能研究 SP1包括其受体鉴定、其检测 具有免疫功能和促进生长的特性。是否存在 将在培养细胞中检测SP1的特异性受体。这个 SP1的潜在免疫抑制作用将被用来研究 刺激淋巴细胞系统。SP1阻断细胞毒T细胞的能力 还将研究细胞作用和免疫球蛋白结合情况。这个 Sp1的生长效应将通过培养细胞进行研究。

The long term objective of this proposal is to understand the functional roles of the different species of "pregnancy-specific beta 1-glycoprotein (SP1)" in human pregnancy with a view to enhance the knowledge of factors which affect the establishment of pregnancy and the well-being of the growing fetus. The immediate goals of this proposal include elucidating the structure of the different SP1 cDNAs and genes, demonstrating the factors regulating gene expression, and defining the potential physiological functions of these proteins. Initially, the nucleotide sequence of the different SP1 CDNAs will be determined. Gene-specific oligonucleotide probes will be synthesized basing on the structural differences among these cDNAs. The different species of human SP1 proteins will be prepares by in vitro expression. Specific monoclonal antibodies (MAb) to each species of SP1 will also be prepared. Factors affecting the expression of RNA dot blotting techniques. Gestational effects will be studied using placental tissues and maternal serum. Tissue specificity of SP1 gene products will be examined in tissue cultures as well as using cDNA libraries derived from different tissues. Effects of hormones and other molecules will be examined using trophoblastic cultures. SP1 genes will be characterized with respect to the intron/exon location as well as the transcription initiation and termination sites using techniques such as restriction enzyme mapping, Northern and Southern blot analysis, S1 nuclease protection assay, primer extension and nuclear run-on assay. Organization of SP1 gene family will be studied by chromosomal localization using gene-specific probes and chromosomal walking. Functional studies of SP1 include identification of its receptor, examination of its immunological functions and growth enhancing properties. Existence of specific receptors for SP1 will be examined in cultured cells. The potential immunosuppressive effect of SP1 will be investigated using stimulated lymphocyte systems. The ability of SP1 in blocking cytotoxic T cell action and in binding immunoglobulins will also be investigated. THe growth effect of Sp1 will be studied using cultured cells.