Molecular Biological Studies on new regulatory factors in the control of blood pressure and body fluid with reference to the pathogenesis of hypertension

D-氨基酸氧化酶（EC 1.4.3.3，DAO）是一种以FAD为辅基的黄素酶，催化多种D-氨基酸的氧化脱氨。据报道，这种酶位于过氧化物酶体的细胞内，DAO被认为是过氧化物酶体的特征性标记酶。人类基因组含有DAO基因的单拷贝，包括11个外显子，跨度为20 kb。通过用用于PCR分析的独特序列引物筛选一组中国仓鼠和人类体细胞杂交体，将人DAO基因座分配到染色体12。在第一个内含子中存在两个交替的嘧啶和嘌呤核苷酸序列（CA）20和（CA）17。为了寻找人类DAO基因中CA重复微卫星DNA周围序列的异质性，分析了21个独立的人类基因组DNA，包括高加索人、黑人和东方人群，发现人类DAO基因是多态性标记的丰富来源。有 ...更多信息 在第一内含子中有三个潜在的遗传多态性序列，即五核苷酸ATTTT和2聚T区。肾素结合蛋白（RnBP）是一种与肾素结合的蛋白质，形成称为高分子量肾素的蛋白质复合物。为探讨RnBP的生理功能，采用北方印迹杂交技术检测了RnBP在大鼠主动脉中的基因表达。RnBP基因在大鼠主动脉、肾脏、肾上腺、脑和卵巢中均有表达。猪主动脉内皮细胞中RnBP基因表达水平较高，为肾脏的40%。为了研究肾素基因表达与RnBP基因表达的关系，采用RT-PCR方法从猪肾mRNA中分离出猪肾素cDNA片段，并测定其核苷酸序列。最后，对猪血管内皮细胞中肾素mRNA的RT-PCR分析表明，在同一内皮细胞中也有肾素基因的表达。在培养的内皮细胞中，RnBP基因表达被Angiotens in II和8-溴cAMP上调。因此，RnBP可能作为血管肾素-血管紧张素系统的一员，在血管内皮参与血压调节。少

D-Amino acid oxidase (EC1.4.3.3, DAO) is a flavoenzyme with FAD as its prosthetic group that catalyzes the oxidative deamination of a wide range of Damino acids. This enzyme was reported to be intracellularly localized in peroxisomes and DAO is regarded as a characteristic marker enzyme of the peroxisomes. The human genome contains a single copy of DAO gene, comprising 11 exons and spanning 20kb. The human DAO locus was assigned to chromosome 12 by screening a panel of Chinese hamster and human somatic cell hybrids with unique sequence primers for PCR analysis. Two sequences of alternating pyrimidine and purine nucleotides, (CA) 20 and (CA) 17, are present in the first intron. In order to search for heterogeneity in the sequences around the CA repeat microsatellite DNA in the human DAO gene, 21 independent human genomic DNAs, comprising Caucasian, Black and Oriental populations were analyzed, and the human DAO gene was found to be an abundant source of polymorphic markers. There are th … More ree other potential sequences for genetic polymorphism in the first intron, namely pentanucleotide ATTTT and 2 poly T tract.Renin-binding protein (RnBP) is a protein that binds to renin, forming a protein complex called high molecular weight renin. In search of its physiological function, gene expression of RnBP in rat aorta were examined by Northern blot hybridization. RnBP gene expression was observed in rat aorta, in addition to kidney, adrenal gland, brain and ovary. The arterial endothelial cells derived from porcine aorta exhibited high level of RnBP gene expression, 40% of the level observed in the kidney. In order to study renin gene expression in relation to that of RnBP,porcine renin cDNA fragment was isolated by RT-PCR analysis of porcine kidney mRNA,followed by determination of the nucleotide sequence for porcine renin. Finally, the RT-PCR analysis of porcine renin mRNA in endothelial cells has shown that renin gene is also expressed in the same endothelium. RnBP gene expression was shown to be up-regulated by Angiotens in II and 8-bromo cAMP in cultured endothelial cells. RnBP,therefore, may participate in blood pressure regulation at vascular endothelium as a member of vascular reninangiotensin system. Less