mCD20-腺病毒载体的构建  

Construction of mCD20-adenovirus vector

作　　者：李慧[1] 刘兵[1] 王友臣[2] 谭晓华[1] 

机构地区：[1]解放军北京军区总医院血液科,北京市100700 [2]解放军第三军医大学研究生队,重庆市400038

出　　处：《中国组织工程研究与临床康复》2007年第37期7405-7408,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：目的:构建编码鼠CD20(mCD20)基因的腺病毒载体,即AdmCD20重组质粒,为后续将其感染DC作为疫苗免疫小鼠做准备工作,为CD20基因治疗淋巴瘤的研究奠定基础。方法:实验于2006-03/2007-04在解放军北京军区总医院血液科实验室完成。运用分子生物学技术,利用本实验室保存的mCD20,以PCR凝胶电泳、酶切进行鉴定后,将该基因cDNA片段插入腺病毒穿梭质粒pDC315中,转化至高效感受态DH5a细菌细胞中进行扩增,提取质粒,获得重组质粒腺病毒穿梭质粒pDC315-mCD20,然后采用Ad MaxTM腺病毒载体包装体系,构建Ad5/35mCD20。结果:顺序成功重组pcDNA3.1-mCD20质粒,腺病毒穿梭质粒pDC315-mCD20,然后采用Ad MaxTM腺病毒载体包装体系成功构建编码鼠CD20基因的腺病毒载体。结论:表达mCD20基因的腺病毒载体构建成功,此方法准确可行,为后续实验奠定基础。AIM： To construct recombinant plasmid that adenovirus encoding the mouse CD20 gene （mCD20）-AdmCD20, lay a primary foundation for that DC were infected by Ad5/35 mCD20 and further study of CD20 gene therapy of lymphoma. METHODS： The experiment was conducted at the Laboratory of Department of Hematopathy of Beijing General Hospital of Chinese PLA from March 2006 to April 2007. The fragment of mCD20 in this laboratory was inserted into adenovirus shuttle plasmid pDC315 after being identified with PCR, restriction enzyme digesting and DNA sequencing by molecular biology technology. The recombinant plasmid were transformed into the cells of DH5abacteria, then the plasmids were extracted from the amplification, and there was the recombinant plasmid pDC315-mCD20, and then Ad5/35 mCD20 was constructed by Ad MaxTM adenovirus vector packaging system. RESULTS： The recombinant plasmid pcDNA3.1-mCD20 and pDC315-mCD20 were successfully constructed, and then Ad5/35 mCD20 was successful constructed by Ad MaxTM adenovirus vector packaging system. CONCLUSION： The successful expression of recombinant plasmid Ad5/35 mCD20 is feasible and exact, and this way would be helpful to the following study.

关 键 词：腺病毒 穿梭质粒 CD20 重组质粒 基因治疗 

分 类 号：R733.4[医药卫生—肿瘤]