| 靶向人VEGF165的SiRNA慢病毒的构建和鉴定 |
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| 引用本文： | 刘蓓,马戈甲,宋保强. 靶向人VEGF165的SiRNA慢病毒的构建和鉴定[J]. 中国美容医学, 2011, 20(7): 1084-1086 |
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| 作者姓名： | 刘蓓 马戈甲 宋保强 |
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| 作者单位： | 第四军医大学西京医院全军整形外科研究所,陕西西安,710032 |
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| 基金项目： | 国家自然科学基金(30870982) |
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| 摘 要： | 目的：构建靶向VEGF基因的SiRNA慢病毒载体,并包装病毒。方法：针对已经筛选确定的SiRNA有效靶序列,合成靶序列的OligoDNA,经双酶切后与PGC SIL-GFP慢病毒载体连接,构建慢病毒载体PGCSIL-SiVEGF,以293T细胞包装得到干扰病毒LV-SiVEGF,根据细胞表达绿色荧光蛋白水平计算病毒滴度;病毒感染人脐静脉内皮细胞HUVEC,通过定量RT-PCR检测VEGF的mRNA表达。结果：测序结果表明,靶向人VEGF的SiRNA慢病毒载体构建成功,包装后的慢病毒滴度为2×109 ifu/μl;感染HUVEC细胞后,实验组和对照组相比VEGF mRNA表达水平显著降低。结论：成功构建人VEGF SiRNA慢病毒LV-SiVEGF。
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| 关 键 词： | VEGF 慢病毒载体 RNA干扰技术 |
Construction and identification of lentivirus of RNA interference targeting VEGF165 gene |
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| LIU Bei,MA Ge-jia,SONG Bao-qiang. Construction and identification of lentivirus of RNA interference targeting VEGF165 gene[J]. Chinese Journal of Aesthetic Medicine, 2011, 20(7): 1084-1086 |
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| Authors: | LIU Bei MA Ge-jia SONG Bao-qiang |
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| Affiliation: | LIU Bei,MA Ge-jia,SONG Bao-qiang(Department of Plastic Surgery,Xijing Hospital,The Fourth Military Medical University,Xi'an,710032 Shaanxi,China) |
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| Abstract: | Objective To construct a lentiviral vector of RNA interfence(siRNA) of VEGF165 gene,and preparing mature virus.Methods The effective sequence of siRNA targeting VEGF165 was confirmed in our previous study.The complementary DNA containing both sense and antisense Oligo DNA of the targeting sequence was synthesized and cloned into the PGC SIL-GFP lentivector obtaining the VEGF165 siRNA virus vector named pGCSIL-SiVEGF.293T cells were cotransfected with pGCSIL-SiVEGF and lentivirus packaging plasmids mixtures ... |
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| Keywords: | VEGF lentivirus RNA interference |
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