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目的检测大鼠CTLA4-FasL重组腺相关病毒载体感染大鼠关节炎性成纤维样滑膜细胞后目的基因的表达。方法构建大鼠CTLA4-FasL融合基因重组腺相关病毒载体,制备重组病毒,体外感染从佐剂诱导关节炎的大鼠关节中分离的炎性成纤维样滑膜细胞,利用Flag标签纯化目的蛋白,ELISA和Western blot等方法检测目的蛋白的表达。结果获得含有大鼠CTLA4-FasL融合基因的重组腺相关病毒,感染炎性成纤维样滑膜细胞后能够分泌性表达目的蛋白。结论构建并制备的CTLA4-FasL重组腺相关病毒能够有效感染炎性成纤维样滑膜细胞并促使期表达CTLA4-FasL融合蛋白,为今后关节炎治疗的体内实验研究奠定了基础。
Objective To detect the expression of the target gene after the rat CTLA4-FasL recombinant adeno-associated virus vector was infected into arthritis fibroblast-like synoviocytes. Methods The recombinant adeno-associated virus vector of CTLA4-FasL fusion gene was constructed and the recombinant virus was prepared. The inflammatory fibroblast-like synoviocytes isolated from the joints of adjuvant-induced arthritis rats were infected in vitro. The target protein was purified by Flag tag, ELISA and Western blot and other methods to detect the expression of the target protein. Results The recombinant adeno-associated virus (CTLA4-FasL) containing rat CTLA4-FasL fusion gene was obtained and secreted the target protein after infection with inflammatory fibroblast-like synoviocytes. Conclusion CTLA4-FasL recombinant adeno-associated virus constructed and prepared can effectively infect inflammatory fibroblast-like synoviocytes and promote the expression of CTLA4-FasL fusion protein in the future, which lays the foundation for future in vivo experimental research on the treatment of arthritis.