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| 棕点湍蛙皮肤分泌液中促胰岛素释放肽的分离纯化、基因克隆及活性分析 | |
| 引用本文： | 冯浩,李宗杰,吕毅,王美娟,李建许.棕点湍蛙皮肤分泌液中促胰岛素释放肽的分离纯化、基因克隆及活性分析[J].动物学研究,2009,30(2):165-170. |
| 作者姓名： | 冯浩 李宗杰 吕毅 王美娟 李建许 |
| 作者单位： | 1. 南京农业大学生命科学学院农业部农业环境微生物工程重点开放实验室,江苏,南京,210095: 2. 上海应用技术学院,上海,200235 3. 中国科学院昆明动物研究所,动物模型和人类疾病机理重点实验室,云南,昆明,650223 |
| 摘 要： | 通过分离纯化棕点湍蛙(Amolops loloensis)皮肤分泌液中的生物活性物质,得到有促胰岛素释放活性的分离峰,并鉴定其结构.采用葡聚糖Sephadex G-50凝胶层析和反相高效液相(RP-HPLC)等手段对棕点湍蛙皮肤分泌液进行分离纯化,利用胰岛素释放实验进行活性检测,Edman降解法测定活性峰的氨基酸序列,反转录法构建cDNA文库并克隆其基因.得到一个具有显著的促胰岛素释放活性的十六肽,测得其氨基酸序列为:FMPIvGKsMSGLSGKL-NH2,命名为amolopin-1.由cDNA(开放阅读框为192bp)推导的氨基酸一级结构显示,其前体由64个氨基酸残基(aa)组成,包括高度保守的信号肽(22aa),酸性肽以及成熟肽.经过数据库序列比对,从棕点湍蛙皮肤中得到一个新的促胰岛素释放肽,进一步分析其作用机理和药代动力学,极有可能得到一个新的治疗糖尿病的降糖药物. |
| 关 键 词： | 棕点湍蛙 皮肤分泌液 促胰岛素释放肽 胰岛素分泌 |
| 收稿时间： | 2008-10-16 |
Isolation and Molecular Cloning of Insulinotropic Peptide from the Skin Secretions of Amolops loloensis and the Study of Its Bioactivities |
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| FENG Hao,LI Zong-jie,LV Yi,WANG Mei-juan,LI Jian-xu.Isolation and Molecular Cloning of Insulinotropic Peptide from the Skin Secretions of Amolops loloensis and the Study of Its Bioactivities[J].Zoological Research,2009,30(2):165-170. | |
| Authors: | FENG Hao LI Zong-jie LV Yi WANG Mei-juan LI Jian-xu |
| Institution: | 1. Key Laboratory of Microbiological Engineering of Agricultural Environment;Ministry of Agriculture;Life Sciences College of Nanjing Agricultural University;Nanjing 210095;China;2. Shanghai Institute of Technology;Shanghai 200235;3. Biotoxin Units of Key Laboratory of Animal Models and Human Disease Mechanisms;Kunming Institute of Zoology;the Chinese Academy of Sciences;Kunming 650223;China |
| Abstract: | To investigate the bioactive component from the skin secretion of Amolops loloensis, isolated peaks with insulin-releasing activity were purified and structurally determined. Skin secretions were isolated by Gel filtration and Reversed phase high-performance liquid chromatography, and tested with insulin-releasing assay. The amino acid sequence of bioactive peak was determined by Edman degradation and identified by gene clone method. A 16-amino peptide with obvious insulin-releasing activity was obtained an... |
| Keywords: | Amolops loloensis Skin secretion Insulinotropic peptide Insulin release |
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