CLONING RHESUS CD94 NKG2 A & NKG2 C NK CELL RECEPTOR CDNAS FROM PREGNANT DECIDU

目的确定Sp家族成员结合位点的作用 生长激素变异基因转录。结果以确定 普遍存在的Sp家族转录因子在GH-V中的作用 转录，我们进行了共转染实验，在其中 将不同数量的Sp1、Sp3或两种因子导入细胞 以确定这些因子是否可以直接激活转录 发起人。SP1处理诱导了基础和基础的 和cAMP刺激的转录；SP3没有刺激作用 基础活性或cAMP诱导的活性，并可能在 更高的水平。两者之间没有协同作用。 各种因素。这些结果表明Sp./3要么不起作用，要么 对GH-V转录没有限制。这位推动者的一个关键事实 Sp位点和邻近元素是转录所必需的。 我们认为，由于转录的基本要素 激活位于-140/-111和-65/-42区域内， 在这两个因素之间存在着合作活动 地区。我们希望确定是否有严格的 这两个元素的要求，或者这两个元素中的任何一个 可以同时取代另一个，从而提供了一个可能的 酵母增强剂1-杂交筛选方法。构造是 使用-140/-105或-75/-35串连器(多个前向和 相反的安排)。瞬时转染法的结果 实验证明，两种药物具有协同作用。 添加1或2个额外的-140/-105元件拷贝，以及 有要求复印件必须在同一张纸上 定向(即，头对头排列不起作用 而头到尾的排列在 单份)。因此，结果表明1)多个副本 激活所必需的元素，2)特定 空间组织是协同激活的必要条件；3) 分离的元件在异源启动子的作用下是有效的。钥匙 胎盘，基因转录，胎盘生长激素， 转录因子资助NIH R01 HD26458

OBJECTIVE To determine the roles of Sp family member binding sites in growth hormone variant gene transcription. RESULTS To determine the role ubiquitous Sp famila transcription factors may play in GH-V transcription, we conducted co-transfection experiments in which varied amounts of Sp1, Sp3 or both factors were introduced into cells to determine if the factors can directly activate transcription from the promoter. Sp1 treatment induced a modest increase in both basal and cAMP-stimulated transcription; Sp3 had no stimulatory effect on either basal or cAMP-induced activity, and may have been inhibitory at higher levels. There was no synergistic activity between the two factors. These results suggest that Sp./3 either plays no role in, or are not limiting for GH-V transcription. A key fact of this promoter is that Sp sites and adjacent elements are required for transcription. We reasoned that since essential elements for transcriptional activation were located within both the -140/-111 and -65/-42 regions, that there was cooperative activity between factors bound in these two regions. We wished to determine whether there was a strict requirement for these two elements, or whether either of the elements could substitute for the other in tandem and thus provide a possible enhancer for yeast 1-hybrid screening approaches. Constructs were prepared with -140/-105 or -75/-35 concatamers (multiple forward and reverse arrangements). The results of transient transfection experiments demonstrated that there was a synergistic effect of addition of 1 or two additional copies of the -140/-105 element, and that there was a requirement for the copies to be in the same orientation (i.e., a head to head arrangement was not functional whereas a head to tail arrangement showed synergistic activity over a single copy). The results thus demonstrate that 1) multiple copies of ea ch of the elements are necessary for activation, 2) specific spatial organization is necessary for cooperative activation, and 3) isolated elements are functional with a heterologous promoter. KEY WORDS placenta, gene transcription, placental growth hormone, transcription factors FUNDING NIH R01 HD26458