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| Dll4基因靶向RNAi重组腺相关病毒载体的构建和高滴度病毒制备 | |
| 引用本文： | 吕伟,陈凛,卫勃,赵云山,李涛,彭正,唐云,李荣. Dll4基因靶向RNAi重组腺相关病毒载体的构建和高滴度病毒制备[J]. 军医进修学院学报, 2009, 30(3): 354-356 |
| 作者姓名： | 吕伟 陈凛 卫勃 赵云山 李涛 彭正 唐云 李荣 |
| 作者单位： | 解放军总医院普通外科,北京,100853 |
| 基金项目： | 国家自然科学基金资助项目(30872468)~~ |
| 摘 要： | 目的：构建一个表达肿瘤血管生成调节基因Delta-like ligand4（Dll4）小干扰RNA的重组腺相关病毒载体,制备靶向性抑制Dll4表达的高滴度重组腺相关病毒。方法：将包含肿瘤血管生成调节基因Dll4特异性小干扰RNA片段的质粒用EcoRI＋SalI双酶切,回收目的片段,将pSNAV2.0质粒用EcoRI＋SalI双酶切后同目的片段连接,脂质体法转染BHK-2l细胞,G418筛选后以辅助病毒感染获取病毒。结果：PCR和测序证实,成功构建包含U6启动子和肿瘤血管生成调节基因Dll4特异性RNA干扰片段的重组腺相关病毒载体,并制备出滴度为2.4×10^11 vectorgenome/L高滴度腺相关病毒。结论：成功构建携带Dll4基因短发夹状干扰RNA的腺相关病毒载体。为下一步探索抗肿瘤血管生成基因治疗的新途径提供实验基础。 |
| 关 键 词： | Delta-likeligand4(Dll4) RNA干扰 重组腺相关病毒 |
Construction of recombined adeno-associated virus vector expressing Dll4 short RNA interference and preparation of high-titer virus |
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| LV Wei,CHEN Lin,WEI Bo,ZHAO Yun-shan,LI Tao,PENG Zheng,TANG Yun,LI Rong. Construction of recombined adeno-associated virus vector expressing Dll4 short RNA interference and preparation of high-titer virus[J]. Academic Journal of Pla Postgraduate Medical School, 2009, 30(3): 354-356 | |
| Authors: | LV Wei CHEN Lin WEI Bo ZHAO Yun-shan LI Tao PENG Zheng TANG Yun LI Rong |
| Affiliation: | Department of General Surgery;Chinese PLA General Hospital;Beijing 100853;China |
| Abstract: | Objective:To construct the recombinant adeno-associated virus vector expressing tumor angiogenesis-related gene Dll4 short RNA interference(RNAi) and to use it for the preparation of high-titer virus.Methods:pDC316-Dll4-shRNA plasmid,constructed with Dll4 siRNA,was digested with EcoRI and SalI.A 1 500bp fragment,containing U6 promoter and Dll4 siRNA,was obtained and inserted into the adeno-associated virus vector plasmid pSNAV2.0 digested with EcoRI+SalI.Positive vectors were analyzed through enzyme digesti... |
| Keywords: | delta-like ligand 4 （Dll4） RNA interference recombinant adeno-associated virus （rAAV） |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |