基础医学与临床 ›› 2018, Vol. 38 ›› Issue (3): 335-339.

• 研究论文 • 上一篇    下一篇

抗小鼠移码因子1蛋白多抗制备及其在脂肪细胞分化中的表达

王珺怡1,朱建强1,贾德政1,袁新1,帕孜丽耶·亚森1,关亚群2,梁小弟1   

  1. 1. 新疆医科大学
    2. 新疆医科大学基础医学院生物化学与分子生物学教研室
  • 收稿日期:2017-05-12 修回日期:2017-12-08 出版日期:2018-03-05 发布日期:2018-02-27
  • 通讯作者: 梁小弟 E-mail:xiaodiliang@xjmu.edu.cn
  • 基金资助:
    新疆医科大学科研创新基金;国家大学生创新性实验计划项目

Preparation of antibody against mouse UPF1 and the expression of UPF1 in adipose cell differentiation

  • Received:2017-05-12 Revised:2017-12-08 Online:2018-03-05 Published:2018-02-27

摘要: 目的 制备小鼠上游移码因子1(UPF1)蛋白多克隆抗体并探讨脂肪细胞分化过程中UPF1蛋白的表达。方法 构建UPF1蛋白表达载体,制备并纯化兔抗小鼠UPF1抗体;诱导3T3-L1细胞分化,并用免疫共沉淀(CoIP)观察脂肪细胞分化过程中UPF1蛋白的表达。结果 1)制备了效价≥64万且与重组mUPF1蛋白和天然mUPF1蛋白均有很强的特异性结合能力的兔抗小鼠UPF1的多克隆抗体;2)小鼠3T3-L1细胞成脂诱导过程中UPF1蛋白表达量未发生明显变化;3)脂肪细胞分化过程中UPF1蛋白与UPF2蛋白相互作用增加。结论 1)以mUPF1蛋白C端550个氨基酸为抗原制备的抗体能够有效识别mUPF1蛋白;2)在脂肪细胞分化过程中UPF1蛋白与UPF2蛋白互作增强。

关键词: 脂肪细胞分化, UPF1, 无义介导的mRNA降解, 抗体制备, 肥胖

Abstract: Objective To prepare polyclonal antibodies against mouse UPF1 protein and to investigate the expression of UPF1 protein during adipocyte differentiation. Methods UPF1 protein expression vector was constructed to prepare and purify rabbit UPF1 antibody. The differentation of 3T3-L1 cells were induced and to the expression of UPF1 was detected by CoIP. Results 1) High specific mUPF1 polyclonal antibody was prepared and the titer of this antibody reached 640000; 2) The expression of UPF1 protein does not change during adipogenesis; 3) In the process of adipocyte differentiation , interaction of UPF1 and UPF2 is increased. Conclusions 1) The polyclonal antibodies prepared by using 550 amino acids at the C terminal of mUPF1 protein could effectively recognize intact mUPF1 protein; 2)The interaction of UPF1 protein with UPF2 protein during adipocyte differentiation was enhanced.

Key words: Adipogenesis, UPF1, NMD, Antibody, Obesity

中图分类号: