逆转录病毒载体PLXRN介导的人IL-1Ra在关节滑膜细胞中的表达  

Construction of a Recombinant Retroviral Vector Containing Human IL-1Ra cDNA and Its Expression in Synoviocytes

作　　者：张晓玲[1] 于长隆[2] 毛泽斌[3] 傅欣[2] 张继英[2] 

机构地区：[1]山西大学体育学院,山西太原030006 [2]北京大学第三医院运动医学研究所,北京100083 [3]北京大学医学部生物化学与分子生物学系,北京100083

出　　处：《体育科学》2005年第7期21-24,30,共5页China Sport Science

基　　金：国家科技攻关计划课题(2001BA904B02);高等学校校内人文社会科学研究基金项目(0409003)

摘　　要：目的:建立逆转录病毒介导的人IL-1Ra体外表达体系。方法:应用DNA重组技术,将人IL-1RacDNA片段重组到逆转录病毒质粒pLXRN中,经PT67细胞包装后,产生的重组逆转录病毒感染原代关节滑膜细胞,用ELISA检测IL-1Ra表达。结果:重组pLXRN-IL-1Ra质粒,经酶切鉴定正确。重组逆转录病毒pLXRN-IL-1Ra滴度可达5.5×104CFU/ml,感染原代关节滑膜细胞后能稳定表达。结论:逆转录病毒能介导人IL-1Ra在关节滑膜细胞中的稳定表达,为下一步开展基因治疗运动创伤性骨关节炎奠定了基础。To construct a retroviral mediated expression system of human interleukin-1 receptor-antagonist protein (IL-1Ra).Recombinant retroviral vector pLXRN-IL-1Ra was generated by cloning a full length human IL-1Ra cDNA into retroviral vector pLXRN.Synoviocytes were infected with the viral supernatant from the highest productive PT 67 clones.ELISA was used for detection of IL-1Ra.The restriction enzyme analysis with EcoR 1 showed that the recombinant retroviral vector had been constructed correctly.The titer assayed on NIH3T3 cells was up to 5.5 ×10~4CFU/ml.Using the ELISA method,the IL-1Ra protein was expressed in synoviocytes after pLXRN-IL-1Ra infection.It concluded that the constructed retroviral vector was able to generate effective expression of human IL-1Ra in synoviocytes,with potential utility in the gene therapy for osteoarthritis in sports medicine.

关 键 词：IL-1RA 逆转录病毒载体 基因转移 创伤性骨关节炎 

分 类 号：G804.5[文化科学—运动人体科学]