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

克隆恒河猴 CD94 NKG2 A

• 批准号: 6312971
• 负责人: THADDEUS G GOLOS
• 金额: $ 5.44万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1976
• 资助国家: 美国
• 起止时间: 1976-06-01 至 2002-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6312971
• 关键词: animal tissue; biotechnology; endocrine gland /system; genome; hormones; mother /infant health care; reproductive system

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

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 确定 普遍存在的 Sp 家族转录因子在 GH-V 中可能发挥的作用 转录，我们进行了共转染实验，其中 将不同量的 Sp1、Sp3 或两种因子引入细胞中 以确定这些因子是否可以直接激活转录 发起人。 Sp1 处理诱导了基础值的适度增加 和 cAMP 刺激的转录； Sp3没有刺激作用 基础或 cAMP 诱导的活性，并且可能在 更高的水平。 两者之间不存在协同活性 因素。 这些结果表明 Sp./3 要么不发挥作用，要么 不限制 GH-V 转录。 该发起人的一个关键事实 是Sp位点和邻近元件是转录所必需的。 我们推断，由于转录的必需元件 激活位于-140/-111和-65/-42区域内， 这两个因素之间存在合作活动 地区。 我们希望确定是否存在严格的 对这两个要素的要求，或者其中一个要素是否 可以同时替代另一个，从而提供一种可能的 酵母 1-杂交筛选方法的增强剂。 构造是 用-140/-105或-75/-35串联体制备（多个正向和 反向安排）。 瞬时转染结果 实验证明，两者之间存在协同效应 添加 1 或两个额外的 -140/-105 元素副本，以及 要求副本必须在同一位置 方向（即，头对头的安排不起作用 而头尾排列显示出协同活性 单份）。 因此，结果表明 1) 多个副本 每个元素都是激活所必需的，2) 特定的 空间组织对于合作激活是必要的，并且3） 分离的元件与异源启动子一起发挥功能。 钥匙 WORDS 胎盘、基因转录、胎盘生长激素、 转录因子 资助 NIH R01 HD26458