GENETIC STUDIES OF PREGNANCY-SPECIFIC B1 GLYCOPROTEIN

本提案的长期目标是了解 不同种类的“妊娠特异性β 1-糖蛋白”的作用 (SP1)“在人类怀孕，以期提高知识的因素 影响怀孕的建立和妇女的福祉， 胎儿发育 该提案的近期目标包括阐明 不同SP1 cDNA和基因的结构，证明了 调节基因表达的因素，并确定潜在的 这些蛋白质的生理功能。 最初，核苷酸 将确定不同SP1 cDNA的序列。 基因特异 寡核苷酸探针将基于结构合成。 这些cDNA之间的差异。 不同种类的人SP1蛋白 将通过体外表达制备。特异性单克隆抗体 (MAb)也将制备SP1的每种物种的样品。影响因素 RNA斑点印迹技术。 地球引力效应将是 使用胎盘组织和母体血清进行研究。 的组织特异性 将在组织培养物中以及使用cDNA检测SP1基因产物 来自不同组织的文库。 激素和其他的影响 将使用滋养层培养物检测分子。 SP1基因将 特征在于内含子/外显子位置以及 转录起始和终止位点， 限制性内切酶图谱，北方和南方印迹分析，S1 核酸酶保护测定、引物延伸和核连续测定。 SP1基因家族的组织结构将通过染色体定位进行研究 使用基因特异性探针和染色体步移。 的功能研究 SP1包括其受体的鉴定、其受体的检查 免疫功能和生长促进特性。 存在 将在培养的细胞中检测SP1的特异性受体。 的 SP1的潜在免疫抑制作用将使用 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结合免疫球蛋白。 的 将使用培养的细胞研究Sp1的生长效果。

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.