STRUCTURE/FUNCTION OF A PREGNANCY-ASSOCIATED PROTEIN

怀孕相关蛋白质的结构/功能

• 批准号: 2205582
• 负责人: THOMAS ROSS HANSEN
• 金额: $ 8.29万
• 依托单位: UNIVERSITY OF WYOMING
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-12-01 至 1999-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2205582
• 关键词: antiserum; cell type; complementary DNA; computer assisted sequence analysis; cow; endometrium; gene expression; genetic regulation; genetic transcription; high performance liquid chromatography; immunoaffinity chromatography; in situ hybridization; laboratory rabbit; northern blottings; pregnancy; protein purification; protein sequence; protein structure function; proteins; sex cycle; southern blotting; species difference; synthetic peptide; tissue /cell culture; western blottings

The long-term objectives of the research ar to understand the function, regulation, and mechanism of action of a 16 kDa protein that is secreted in high amounts during pregnancy by the bovine uterine endometrium. The function of this protein has implications in several mammalian species, including humans, since it shares epitopes with ubiquitin and a 15-kDa ubiquitin cross-reactive protein (UCRP), and is regulated by conceptus- derived interferon tau (IFN-). Thus, this protein may serve as an index of early pregnancy and potentially, fertility. To meet the long-term objectives of the research, we intend to focus on the elucidation of structure and pregnancy-associated expression of the 16-kDa protein. Specifically, amino-terminal and internal peptide amino acid sequence will be determined following purification by using fast phase liquid chromatography. The cDNA(s) encoding the 16-kDa protein will be sequenced and used in Northern blots and in situ hybridization to confirm pregnancy-associated transcription. The number, complexity and distribution of genes across species will be studied by using Southern blots. A more specific polyclonal antiserum will be prepared and used to detect the 16-kDa protein in uterine secretions and serum. It also will be used to make an immunoaffinity matrix that will aid in purification. Immunosuppressive activity will be determined in rosette inhibition and anti-proliferative tests. Future experiments, by using cDNAs to generate recombinant 16-kDa protein, will be directed towards identifying function and mechanism of action during early pregnancy. Study of this 16-kDa protein has potential implication in human fertility, since communication between the conceptus and uterus is similar across many mammalian species.

研究AR的长期目标了解该功能， 调节和16 kDa蛋白的作用机理 牛子宫内膜在怀孕期间的大量。 这 该蛋白的功能对几种哺乳动物物种具有影响， 包括人类，因为它与泛素和15 kDa共享表位 泛素交叉反应蛋白（UCRP），受概念的调节 派生的干扰素tau（IFN-）。 因此，该蛋白可以用作 早期怀孕，可能是生育能力。 为了满足研究的长期目标，我们打算专注于 阐明结构和与妊娠相关的16 kDa表达 蛋白质。 具体而言，氨基末端和内部肽氨基酸 通过使用快速相纯化后，将确定序列 液相色谱。 编码16 kDa蛋白的cDNA将是 测序并用于北印迹和原位杂交以确认 与妊娠相关的转录。 数字，复杂性和 通过使用南部研究，将研究跨物种的基因的分布 印迹。 将准备更具体的多克隆抗血清 检测子宫分泌和血清中的16 kDa蛋白。 也将是 用于制作有助于纯化的免疫亲和力基质。 免疫抑制活性将在玫瑰花塞抑制和 抗增殖测试。 未来的实验，通过使用cDNA生成 重组16-kDa蛋白将针对识别功能 和怀孕初期的作用机制。 研究这个16千克 蛋白质对人类的生育有潜在的影响 在许多哺乳动物物种中，概念和子宫之间相似。