实验性家兔骨关节炎的基因治疗Ⅰ:带有治疗基因的穿梭载体pLXSNhIL-1ra的构建及真核表达

Gene Therapy of Experimental Osteoarthritis in Rabbit. I:Combination and Eucaryotic Expression of Shuttle Vector pLXSNhIL-1ra with Therapy Gene

本研究用含有人白细胞介素 - 1受体拮抗蛋白基因 (hIL - 1ra)的质粒pCD1-hIL - 1a和反转录病毒穿梭质粒pLXSN ,通过基因重组的方法 ,重组完成带有治疗基因hIL - 1ra的反转录病毒穿梭质粒pLXSNhIL - 1ra。通过DNA限制性内切酶的酶切鉴定 ,证明插入方向正确 ,通过把pLXSNhIL - 1ra穿梭质粒转染到Cos - 7细胞中进行真核表达 ,证明该质粒能够成功表达hIL - 1ra基因产物 ,为下一步用hIL - 1ra对骨关节炎动物模型进行基因治疗工作奠定了基础。

Plasmid pCD1-hIL-1a recombined with hIL-1ra gene and retrovirus shuttle plasmid pLXSN were used. By gene recombination technique, a new retrovirus shuttle plasmid pLXSNhIL-1ra recombined with hIL-1ra gene was got. The insertion direction was demonstrated to be correct by DNA restriction endonuclease reaction. The hIL-1ra gene expression was successfully tested by transfecting pLXSNhIL-1ra into Cos-7 cells. The groundwork had been done for next step osteoarthritis gene therapy with hIL-1ra gene by animal model.