摘要
【目的】构建鼠IL 2Rα与β链基因的反义RNA真核表达质粒 ,为反义RNA基因治疗细胞免疫相关疾病作准备。【方法】用限制性内切酶从克隆载体上切下鼠IL 2Rα与β链cDNA及鼠IL 2启动子 ,克隆入真核表达质粒 pcDNA3的多克隆位点上 ,用酶切或PCR法鉴定正反连接。【结果】得到 4个反向连接的重组质粒 :pcAnti mIL 2Rα ,pcAnti mIL 2Rβ,pcAnti mIL 2Rαβ及pciAnti mIL 2Rαβ。前三者由CMV强启动子指导鼠IL 2R基因反义RNA的表达 ,后者由CMV启动子与鼠IL 2靶向可诱导型启动子组成的融合启动子指导反义RNA的表达。【结论】成功构建了 4个不同类型的鼠IL 2R基因反义RNA真核表达质粒。
Objective To construct a series of mouse IL 2R α and β chain gene antisense RNA eukaryotic expression plasmids, to make a path for antisense gene therapy for cellular immunity related diseases. Methods Mouse IL 2R α, β chain cDNA and mouse IL 2 promoter isolated from plasmids were inserted into the multiple cloning site of eukaryotic expression vector pcDNA3. The orientation of ligation was determined by restriction endonucleases or PCR method. Results Four recombinant plasmids were obtained: pcAnti mIL 2Rα, pcAnti mIL 2Rβ, pcAnti mIL 2Rαβ and pciAnti mIL 2Rαβ. The former three plasmids' expression of mouse IL 2R chain gene antisense RNA was controlled by the CMV promoter, and the later one was controlled by the CMV mIL 2 chimeric promoter. Conclusion Four different kinds of mouse IL 2R gene antisense nucleic acid eukaryotic expression plasmids were successfully constructed.
出处
《中山医科大学学报》
CSCD
2000年第2期113-116,共4页
Academic Journal of Sun Yat-sen University of Medical Sciences
基金
广东省自然科学基金!( 963 0 0 3 )
