天津医药

天津医药 ›› 2020, Vol. 48 ›› Issue (8): 711-714.doi: 10.11958/20193697

• 实验研究 • 上一篇    下一篇

真核表达载体介导的ICOSIg基因在大鼠脂肪间充质干细胞中表达的实验研究#br#

耿洁1，2，刘涛3，姜锦4，李光5，黄志伟1，王玉亮2△
  

  1. 1天津医科大学研究生院（邮编300070）；2天津医科大学第二医院检验科，天津市泌尿外科研究所；3天津市第一中心医院器官移植中心，4口腔科；5天津医科大学基础医学院遗传学系
  • 收稿日期:2019-12-09 修回日期:2020-06-23 出版日期:2020-08-15 发布日期:2020-08-12
  • 通讯作者: 王玉亮 E-mail:wang_yu_l@163.com
  • 作者简介:耿洁（1995），女，硕士在读，主要从事移植免疫学相关研究
  • 基金资助:
    国家自然科学基金资助项目（81470982）；国家高技术研究发展计划（863）项目（2012AA021003）

The expression ICOSIg gene mediated by eukaryotic expression vector in rat adipose tissue-derived mesenchymal stem cells#br#

GENG Jie1,2, LIU Tao3, JIANG Jin4, LI Guang5, HUANG Zhi-wei1, WANG Yu-liang2△#br#   

  1. 1 Graduate School of Tianjin Medical University, Tianjin 300070, China; 2 Department of Clinical Laboratory Medicine, the Second Hospital of Tianjin Medical University, Tianjin Institute of Urology; 3 Organ Transplantation Center, 4 Department of Stomatology, Tianjin First Central Hospital; 5 Department of Genetics, School of Basic Medical Sciences, 
    Tianjin Medical University
  • Received:2019-12-09 Revised:2020-06-23 Published:2020-08-15 Online:2020-08-12
  • Contact: WANG Yu-liang E-mail:wang_yu_l@163.com

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摘要： 目的　构建含可诱导共刺激分子（ICOS）融合蛋白（ICOSIg）基因的真核表达载体，探讨其能否在大鼠脂肪间充质干细胞（ADSCs）中表达。方法　克隆编码大鼠ICOS的胞外片段，将其与编码人免疫球蛋白IgG Fc段的基因融合，构建ICOSIg融合基因及其分泌型真核表达载体pcDNA3.1（+）/ICOSIg。经测序鉴定后转染ADSCs，Western blot法检测ICOSIg在ADSCs中的表达。结果　经测序鉴定验证pcDNA3.1（+）/ICOSIg质粒构建成功，且能在ADSCs中成功表达。结论　ICOSIg在ADSCs中成功表达，为进一步研究免疫耐受机制提供了实验基础。

关键词: 间充质基质细胞, 受体, Fc, 基因融合, 质粒, 可诱导共刺激分子, 真核表达载体, 脂肪间充质干细胞

Abstract: Objective　To construct eukaryotic expression vector carrying inducible co-stimulator (ICOS) Ig fusion gene, and investigate the ICOSIg expression in rat adipose tissue-derived mesenchymal stem cells (ADSCs). Methods　The encode of the extracellular domain of rat ICOS was cloned. The encode of the domain was fused with the gene of Fc     region encoding human immunoglobulin IgG, in order to build the ICOSIg fusion gene and the secreted eukaryotic expression vector pcDNA3.1(＋)/ICOSIg. After DNA sequence analysis, the recombinant plasmid was transfected into ADSCs, and the expression of ICOSIg was confirmed by Western blot analysis. Results　The sequencing confirmed that   pcDNA3.1(＋)/ICOSIg plasmid were successfully constructed, and ICOSIg was successfully expressed in ADSCs. Conclusion　The ICOSIg can be successfully expressed in ADSCs, which provides the experimental basis for further research on immune tolerance mechanism.

Key words: mesenchymal stromal cells, receptors, Fc, gene fusion, plasmids, inducible co-stimulator, eukaryotic expression vector, adipose tissue-derived mesenchymal stem cells

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