大鼠CTLA4-FasL融合基因重组腺相关病毒载体的构建、制备及表达  

Construction,preparation and expression of rat CTLA4-FasL fusion gene in recombinant adeno-associated virus

作　　者：王博[1,2] 王芳[2] 张瑾[2] 于继云[2] 张巍[2] 

机构地区：[1]吉林大学畜牧兽医学院,长春130062 [2]军事医学科学院基础医学研究所转化医学研究室,北京100850

出　　处：《免疫学杂志》2011年第12期1078-1082,1085,共6页Immunological Journal

摘　　要：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。We aim to detect the expression of rat CTLA4-FasL fusion gene on arthritic fibroblast-like synoviocytes（FLSs） of recombinant adeno-associated virus（rAAV）-infected rat.The rAAV vector with rat CTLA4-FasL fusion gene was constructed,and then the recombinant virus（rAAV.CTLA4-FasL） was prepared and obtained.The inflammatory FLSs isolated from rat adjuvant-induced arthritic joints were infected with rAAV.CTLA4-FasL and the expressions of target proteins in the supernatants were identified by ELISA and Western after purification with Flag-tag.The results indicated that the inflammatory FLSs infected with rAAV.CTLA4-FasL could secretly express the fusion proteins.We conclude that the rAAV.CTLA4-FasL prepared in this study could effectively infect arthritic FLSs and promote the cell to express CTLA4-FasL fusion proteins,which lay the foundations for future research of therapy for arthritis in vivo experiments.

关 键 词：重组腺相关病毒 CTLA4-FASL 成纤维样滑膜细胞 表达 

分 类 号：R593.22[医药卫生—内科学]