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#

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。

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

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.