人白介素10逆转录病毒表达载体的构建

Construction of retroviral vector carrying human interleukin-10

目的构建一个人白介素10(hIL10)逆转录病毒表达载体,为研究hIL10的生物学活性打下基础。方法用限制性内切酶消化提取质粒pCDHIL10中hIL10全长cDNA,采用定向克隆的方法将hIL10基因插入逆转录病毒载体pLXSN的多克隆位点中,经转化、扩增、限制性内切酶消化,电泳鉴定分析。结果外源性hIL10基因成功插入到逆转录病毒载体pLXSN的中。结论成功完成了人白介素10(hIL10)逆转录病毒表达载体的构建,为今后进一步研究hIL10的生物学活性打下了基础。

Objective To study the effects of hIL-10 on graft rejection, and to set up foundation of its application in immunogene therapy. Retroviral vector carrying hIL-10 cDNA was successfully constructed.Methods To digest pCDHIL-10 with restrictive endonucleases, to isolate and purify hIL-10 complete cDNA from pCDHIL-10 and then subclone cDNA into retroviral vector pLXSN; to identify recombinant retroviral vector pLXHIL-10 by digestion with restrictive endonucleases and Agarose Gel Electrophoresis.Results Exogenous hIL-10 gene was subcloned into retroviral vector pLXSN successfully.Conclusion Retroviral vector carrying hIL-10 cDNA shall lay foundation for researching hIL-10 mechanism and its applications for curing diseases.