QQ客服 官方微博 反馈留言

题名	大蹼铃蟾(Bombina Maxima)皮肤Peroxiredoxin分子克隆及分离纯化
作者	杨洋
学位类别	硕士
答辩日期	2012-11
授予单位	中国科学院大学
授予地点	北京
导师	张云
关键词	大蹼铃蟾 大蹼铃蟾 peroxiredoxin 皮肤分泌物 抗氧化
其他题名	Characterization and Molecular Cloning of Peroxiredoxin From the Bombina Maxima Skin Secretion
学位专业	生物化学与分子生物学
中文摘要	两栖类动物皮肤对其生存具有重要意义，它担负着如呼吸、水分调节、抵抗微生物、抵抗外界物理环境损伤等多种生理功能。目前，在两栖类皮肤中已鉴定出多种具有生物活性的蛋白质和多肽。对于两栖类动物皮肤分泌物中的生物活性成分的研究，会有助于我们更好的探索和理解两栖类动物皮肤的生理功能。通过三步分离纯化过程：DEAE-Sephadex A-50离子交换，Sephadex G-100凝胶过滤和Sephadex G-50凝胶过滤，我们从大蹼铃蟾(Bombina Maxima)皮肤分泌物中分离得到纯化的大蹼铃蟾Peroxiredoxin V（BMS-Prdx VI）。SDS-PAGE电泳显示，该蛋白为单链蛋白质，在还原状态和非还原状态下下表观分子量为26 kDa。经Western Blot方法鉴定和胰酶消化进行蛋白测序，得到两条内肽序列，进行Blast序列比对，确定是BMS-Prdx VI蛋白。进行活性检测发现，BMS-Prdx VI具有谷胱甘肽过氧化物酶的活性。通过大蹼铃蟾(Bombina Maxima)转录组测序，我们得到peroxiredoxin家族中BMS-Prdx I、BMS-Prdx V、BMS-Prdx VI的部分序列。在此基础上，采用cDNA末端快速扩增技术（RACE），得到BMS-Prdx VI cDNA全序列，以及BMS-Prdx I和BMS-Prdx V的成熟肽部分的cDNA序列。根据BMS-Prdx VI序列设计引物，进行实时荧光定量，对大蹼铃蟾心、肝、脾、肺、肾、眼、脑、皮肤进行检测发现，皮肤BMS-Prdx VI的表达量是其他组织的4倍以上，提示其在皮肤中可能有重要的生理作用。
英文摘要	Amphibian skin is a morphologically, biochemically and physiologically complex organ that fulfills a wide range of functions necessary for amphibian survival. A serious functions takes place in the skin, including respiration, water regulation, antimicrobial defense, mechanic damage defense, etc. A number of diverse bioactive components have been isolated and characterized from the amphibian skin secretion and would be contribute to the searching and recognition of physiological function in the amphibian skin.Following the trypsin degeneration, two EP sequences of peroxiredoxin VI were sequencing and provided an evidence of BMS –Prdx VI from the skin secretion .With the identification of this protein by Western Blot，BMS-Prdx VI was purified and characterized form frog Bombina Maxima skin secretion by three steps protocol including DEAE-Sephadex A-50 ion exchange，Sephadex G-100，Sephadex G-50 gel filtration. BMS-Prdx VI is composed of a single peptide chain demonstrated by SDS-PAGE. Under reducing conditions and non-reducing conditions, its apparent molecular weight is 26 kDa. Glutathione peroxidase activity assay revealed that the BMS-Prdx VI would have the antioxidative activity in the skin.According to the data of the Bombina Maxima transcriptome, the full cDNA sequence of the BMS –Prdx VI and muture peptide of BMS –Prdx I and BMS –Prdx V have been cloned. Following design of the primer of BMS Prdx VI for the qRT-PCR, the distribution of the protein in eight tissues and the relative amount have been assayed. The amount of BMS Prdx VI in skin is five times more than that in other seven tissues and this suggests that BMS Prdx VI may play some physical functions in the skin.
语种	中文
公开日期	2012-12-10
内容类型	学位论文
源URL	[http://159.226.149.42:8088/handle/152453/7131]
专题	昆明动物研究所_动物活性蛋白多肽组学
推荐引用方式 GB/T 7714	杨洋. 大蹼铃蟾(Bombina Maxima)皮肤Peroxiredoxin分子克隆及分离纯化[D]. 北京. 中国科学院大学. 2012.

个性服务

查看访问统计

相关权益政策

暂无数据

收藏/分享

除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。