优果科技之MRAP2重组质粒的构建过程.docVIP

南京师范大学中北学院 毕 业 设 计（论 文） （2015届） 题 目： MRAP2重组质粒的构建 专 业： 生物技术 姓 名： 　 陆家明 学 号： 指导教师： 卢山 职 称： 填写日期： 南京师范大学中北学院教务处 制 MRAP2重组质粒的构建 摘要 黑皮质素受体是能量平衡和葡萄糖稳态的关键性调节器，MC1-5 受体能通过各生物系统调节食物摄入、能量消耗、身高以及葡萄糖代谢黑皮质素受体辅助蛋白（MRAP）和它的同系物MRAP2都是GPCR辅助蛋白，在黑皮质素系统中发挥功能。MC4的破坏会导致其严重肥胖和糖尿病。黑皮质素受体（MCRs）是GPCRs的亚族，作为α-、β-、γ-MSH（促黑激素）的受体发挥作用。研究发现，MRAP2在成人肾上腺和大脑中有表达。通过原位杂交技术发现，这个基因似乎主要在大脑的下丘脑腹内测区域表达。MRAP2与MC2R相互作用，MC2R与MRAP2共表达时会形成一个功能性ACTH（促肾上腺皮质激素）受体。MRAP2的表达含量常会升高，而前列腺癌的发生也与肥胖相关联。本研究采用RT-PCR方法，在BPH-1，WPMY-1，LNCaP，22RV1，PC-3和DU145六种前列腺细胞中，发现MRAP2在这些细胞中的表达量依次为：aP。同时，采用重组DNA技术，构建了MRAP2全长重组质粒，为分析该基因在肿瘤中的作用机制，以及阐释肥胖与前列腺肿瘤之间的关系奠定了基础。 关键词：MRAP2，MCRs，重组质粒，前列腺细胞 Abstract Melanocortin receptors are critical modulators of energy balance and glucose homeostasis.MC1-5 receptors regulate food intake, energy expenditure, body length, and glucose metabolism in many biophysiological systems. Melanocortin 2 receptor accessory protein (MRAP), and its homolog, MRAP2, are GPCR accessory proteins that function in the melanocortin system. Disruptions of MC4 receptors result in obesity and diabetes. MCRs are a subfamily of GPCRs that act as the receptors to α-,β-, andγ-MSH. Expression of MRAP2 was identified in human adult adrenal gland and brain. In situ hybridization suggests that this gene appears to be primarily expressed in the ventromedial hypothalamus of the brain. When coexpressed with MRAP2, MC2R forms a functional ACTH receptor. MRAP2 is usually at a high level in obesity cells, and the occurrence and development of the prostate cancer is associated to obesity. In six different prostate cells , the level of expression from high to low is 22RV1，PC-3，DU145，BPH-1，MPMY-1 and LNCaP. We further constructed a recombinant plasmid with recombinant DNA technology, which will be helpful to investigate the functions and mechanisms of MRAP2 in prostate and the relation