表达人白细胞介素10的逆转录病毒载体的构建

Construction of Retroviral Vector Expressing Human Inter Leukin 10

目的 建立一个表达人白细胞介素10(human Interleukin 10,hIL—10)的重组逆转录病毒载体基因转移系统,为下一步的研究工作做准备.方法 用限制性内切酶消化,提取和纯化质粒puhIL—10中hIL全长 cDNA,重组到逆转录病毒载体 pLXSN上,采用斑点杂交筛选,限制性内切酶消化,电泳鉴定分析.结果外源性hIL—10基因成功构建到逆转录病毒载体pLXSN上.结论表达人白细胞介素10的逆转录病毒载体的成功构建,为今后研究hIL—10的作用机制及在治疗疾病中的应用打下了基础.

Object: We have established recombinant retroviral gene transfer system of expressing human Interlerkin 10 for futherstudy. Methods: To digest pUhIL-10 with restrictive endonucleases, to isolate and purify hIL-10 complete cDNA from pUhIL-10 and then subclone hIL-10 cDNA into retroviral vector pLXSN; to identify recombinant retroviral vector pLX(hIL-10) SN by dot blot hybridization, digestion with restrictive endonucleases and Agaro'se Gel Electrophoresis; Results: Exogenous ML-10 gene has been subcloned into retroviral vector pLXSN successfully; Conclusion: Subclone pLX(hIL-10) SN shall lay foundation for researching hIL-10 mechanism and its applications for curing diseases.