Characterizing a novel promoter of mouse MOR-1 gene

描述(由申请人提供)：本申请的主要目标 是为了更好地了解Mu的转录调控 阿片受体基因(Oprm)的分子生物学方法及转基因 技巧。在MOR-1基因被克隆后不久，启动子(EL启动子) 和结构(外显子1-4)。 特色化的。紧随其后的是两种剪接形式的分离， MOR-1A和MOR-IB。进一步说明了MOR-L基因的多样性 我们最近发现了另外9个外显子，12个剪接变体 和一个与新外显子之一相关的新启动子(E11启动子)。这 应用重点在于阐明该基因的详细结构和功能 E11启动子及其与E1启动子的关系。E11启动子是 位于原始e1启动子上游约10kb处。备择 由E11启动子驱动的转录本的剪接产生8个变体 编码一些新的蛋白质以及原始的MOR-1蛋白。 受E11启动子调控的三个变体产生原始的MOR-1 蛋白。因此，这种单一的蛋白质可以由四种剪接变体产生 Mu阿片受体基因受两个不同启动子的控制。 变异mRNAs在不同脑区的差异表达 区域特定的RNA处理。具有核心的神经元启动子活性 已经提出了促进因素和消极因素。具体目的是 实现的总体目标是：(1)识别E11中的顺式作用元件 通过突变分析获得启动子区域；(2)分离DNA结合蛋白 调控启动子活性；(3)E11启动子和 其与转基因小鼠中E1启动子的关系。我的长期目标是 了解MOR-1基因的调控机制并获得 对ITS药理学和生理学意义的认识 监管。E11启动子及其相关变体的意义在于 由反义作图研究建议，其中两个反义探针靶向 外显子11阻断了脊髓吗啡和脊髓上M6G的镇痛作用。《知识》 从应用程序获取的信息将有助于确定复杂性和 MOR-L基因调控的功能重要性、细胞模型的建立和 为研究其基因调控提供了转基因动物模型，并提供了 开发用于疼痛控制和药物的新药的潜在目标 虐待。

DESCRIPTION (provided by the applicant): The primary goal of this application is to obtain a better understanding of transcriptional regulation of the mu opioid receptor gene (Oprm) through molecular biological means and transgenic techniques. Soon after the MOR-1 cDNAs were cloned, the promoter (El promoter) and structure (exons 1-4) of the original MOR-1 gene were identified and characterized. This was soon followed by the isolation of two splice forms, MOR-1A and MOR-IB. The diversity of the MOR-l gene was further illustrated by our recent identification of an additional nine exons, twelve splice variants and a novel promoter associated with one of the new exons (E11promoter). This application focuses on elucidating the detailed structure and function of the E11 promoter and its relationships with E1 promoter. The E11 promoter is located at about10 kb upstream of the original E1 promoter. Alternative splicing of the transcripts driven by the E11 promoter yield eight variants encoding a number of novel proteins as well as the original MOR-1 protein. Three of the variants regulated by the E11 promoter generate the original MOR-1 protein. Thus, this single protein can be generated by four splice variants of the mu opioid receptor gene under the control of two distinct promoters. Differential expression of the variant mRNAs in various brain regions revealed region-specific RNA processing. Neuronal promoter activities with a core promoter and a negative element have been suggested. The specific aims to achieve the overall goal are: (1) Identification of cis-acting elements in E11 promoter region through mutant analysis; (2) Isolation of DNA-binding proteins that regulate promoter activities; (3) Investigation of the E11 promoter and its relationships with E1 promoter in transgenic mice. My long-term goal is to understand the mechanisms by which the MOR-1 gene is regulated and to gain insights into the pharmacological and physiological significance of its regulation. The significance of the E11 promoter and its associated variants is suggested by antisense mapping studies in which two antisense probes targeting exon 11 blocked spinal morphine and supraspinal M6G analgesia. The knowledge obtained from the application will help to determine the complexity and functional importance of MOR-l gene regulation, establish the cell models and the transgenic animal models for studying its gene regulation, and provide potential targets for developing novel drugs useful in pain control and drug abuse.