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Peli1蛋白SUMO修饰位点突变体构建及鉴定

发表时间:2015-06-15  浏览量:1326  下载量:494
全部作者: 李建涛,范敏,王敏,许家艳,杨辛怡,王曼,阙玲t ,李跃华
作者单位: 南京医科大学基础医学院
摘 要: 目的:构建Peli1蛋白小类泛素修饰因子(small ubiquitin-like modifier,SUMO)修饰位点突变体,并进行鉴定。方法:提取C57BL/6J小鼠心肌组织RNA,逆转录成cDNA,采用分子克隆的方法,克隆小鼠Peli1基因,构建真核表达载体;进一步采用点突变的方法,设计特异性引物,构建Peli1 5个SUMO修饰位点的突变体。结果:在构建小鼠Peli1真核表达质粒的基础上,分子测序显示成功构建Peli1 SUMO修饰位点突变体,Western blotting检测突变体蛋白可以体外表达。结论:成功构建Peli1蛋白SUMO修饰位点突变体,为进一步探讨Peli1蛋白功能奠定了基础。
关 键 词: 病理生理学;Peli1蛋白;小类泛素修饰因子修饰;点突变;表达载体
Title: Generation and identification of SUMO modification sites mutation of Peli1 protein
Author: LI Jiantao, FAN Min, WANG Min, XU Jiayan, YANG Xinyi, WANG Man, QUE Lingli, LI Yuehua
Organization: School of Basic Medical Sciences, Nanjing Medical University
Abstract: Objective: To generate and identify small ubiquitin-like modifier (SUMO) modification sites mutation of Peli1 protein. Methods: C57BL/6J mice total mRNA was extracted from mice heart tissue, and cDNA was obtained by reverse transcription. Pelil1 gene was cloned and the eukaryotic expression vector of Peli1 was generated according to the molecule cloning protocols. Furthermore, special mutation primers were designed, and 5 mutations of SUMO modification sites mutation of Peli1 protein was generated. Results: The eukaryotic expression vector of Peli1 was generated, and the mutation vectors of Peli1 were constructed using molecular sequencing and can expressed in vitro according to Western blotting method. Conclusion: The SUMO modification sites mutation of Peli1 protein were successfully generated, which will provide fundamental research ground to explore the function of Peli1 protein.
Key words: pathophysiology; Peli1 protain; small ubiquitin-like modifier modification; site mutation; expression vector
发表期数: 2015年6月第11期
引用格式: 李建涛,范敏,王敏,等. Peli1蛋白SUMO修饰位点突变体构建及鉴定[J]. 中国科技论文在线精品论文,2015,8(11):1147-1151.
 
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