MOLECULAR STUDIES OF MONOAMINE OXIDASES

本项目的总体目标是了解 两种单胺氧化酶(MAO A和MAO B)的分子基础 基因水平。MAO是儿茶酚胺中的一种重要酶 新陈代谢。MAO活性的异常水平在一项 精神障碍的数量。用全身人肝毛 本实验室克隆的A、B两个cDNA的基因组组织 并将研究这些基因的表达情况。这些 无论是基础研究还是临床研究，研究都具有重要意义。 这项为期五年的研究科学家奖的具体目标 应用程序如下所述。使用已建立的细菌 和哺乳动物细胞表达载体，克隆的人 肝脏MAO A和B的cDNA将被表达并催化 将对表达的蛋白质的性质进行表征。 以MAO A和MAO B为探针，进行Northern印迹分析。 这些基因在细胞水平上的表达之间的相关性 人体不同组织中的信使核糖核酸、蛋白质及其催化活性 将会被检查。这项研究将表明组织是否 MAO A和MAO B的特异度控制在 转录或翻译。利用Southern杂交技术分析小麦品种 含有人类染色体的细胞-小鼠DNA杂交以及 人肝MAO的原位杂交定位 A和B基因将被确定。这些基本知识是 对研究MA0基因在精神障碍中的作用至关重要。 将分析MAO A和MAO B的基因组DNA。几个人类 基因组文库(COSMIC和LAMBDA)将使用MAO进行筛选 编码人肝脏MAO A的基因组DNA的A和B基因探针 而B基因将被克隆。基因组的限制酶图谱 然后将构建MAO A和B克隆。更进一步，毛A 和B基因将通过链终止法进行测序，以及 我们将分析它们的启动子区域。 将对MAO A和MAO B的异质性进行研究。毛甲、毛乙 相关的cDNA将从其他人类组织中克隆出来，并且 他们的DNA和氨基酸序列将被比较。因此， 不同组织中MAO A(或B)的异同 将会被清楚地理解。此信息将为您提供 关于用血小板MAO B作为实验指标的有效性的确凿答案 脑MAO B的标志物，并将对 未来的临床研究。

The overall objective of this project is to understand the molecular basis of two types of monoamine oxidase (MAO A and B) at the gene level. MAO is an important enzyme in catecholamine metabolism. Abnormal levels of MAO activity have been shown in a number of mental disorders. With the full-length human liver MAO A and B cDNAs cloned in this laboratory, the genomic organizations and the expressions of these genes will be investigated. These studies have great importance for both basic and clinical research. The specific aims of this five-year Research Scientist Award application are described below. Using the established bacterial and mammalian cell expression plasmid vectors, the cloned human liver MAO A and B cDNAs will be expressed and the catalytic properties of the expressed proteins will be characterized. Using Northern blot analysis with MAO A and B cDNA as probes, the correlation among the expressions of these genes at the levels of mRNA, protein and catalytic activities in various human tissues will be examined. This study will indicate whether the tissue specificity of MAO A and B is controlled at the level of transcription or translation. Using Southern blot analysis of cells containing human chromosomes - mouse DNA hybrid as well as in situ hybridization, the chromosomal location of human liver MAO A and B genes will be determined. This fundamental knowledge is essential for studying the role of MA0 genes in mental disorders. The genomic DNAs For MAO A and B will be analyzed. Several human genomic libraries (cosmic and LAMBDA) will be screened using MAO A and B cDNA probes, the genomic DNAs encoding human liver MAO A and B genes will be cloned. The restriction maps of the genomic MAO A and B clones will then be constructed. Further, the MAO A and B genes will be sequenced by the chain termination method, and their promoter regions will be analyzed. The heterogeneity of MAO A and B will be investigated. MAO A, B and related cDNAs will be cloned from other human tissues, and their DNA and amino acid sequences will be compared. Thus, the similarities and differences of MAO A (or B) from various tissues will be clearly understood. This information will provide a conclusive answer on the validity of using platelet MAO B as a marker for brain MAO B and will have significant implications on future clinical studies.