| 外源性microRNA介导的RNA干扰抑制HBV复制和表达 |
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| 引用本文： | 郜玉峰,余莉,叶珺,邹桂舟,李家斌,沈继龙,李旭.外源性microRNA介导的RNA干扰抑制HBV复制和表达[J].安徽医科大学学报,2010,45(4):443-446. |
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| 作者姓名： | 郜玉峰 余莉 叶珺 邹桂舟 李家斌 沈继龙 李旭 |
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| 作者单位： | 安徽医科大学第二附属医院感染病科,合肥,230601;安徽省人兽共患病实验室,教育部省部共建重点实验室,合肥,230022;安徽医科大学第一附属医院感染病科,合肥,230022 |
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| 摘 要： | 目的 设计并构建靶向HBV的外源性microRNA表达载体,观察其对HBV表达和复制的抑制作用.方法 以HBV P基因和X基因为靶基因,根据pcDNA6.2-G/W-miR载体要求,设计靶向HBV的microRNA序列,进行直接合成获得目的 片段并克隆到载体中,测序正确的载体通过脂质体方法转染HepG2.2.15细胞,RT-PCR、乙肝五项定量和荧光定量PCR检测其对HBV mRNA和HBV DNA的抑制作用.结果 质粒的转染效率为50%～60%,在这种状态下,外源性microRNA表达载体在HBV mRNA和HBsAg、HBeAg和HBV DNA水平上均能显著抑制HepG2.2.15细胞中HBV的复制和表达(P＜0.05).结论 外源性microRNA表达载体构建成功,并能抑制HBV靶基因的表达,为其在慢性HBV感染基因治疗中的应用奠定了基础.
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| 关 键 词： | 肝炎病毒 乙型 RNA 小分子干扰热 |
Inhibition of hepatitis B virus gene replication and expression by artificial microRNA |
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| Gao Yufeng,Yu Li,Ye Jun,et al.Inhibition of hepatitis B virus gene replication and expression by artificial microRNA[J].Acta Universitis Medicinalis Anhui,2010,45(4):443-446. |
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| Authors: | Gao Yufeng Yu Li Ye Jun |
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| Institution: | Gao Yufeng1,Yu Li2,Ye Jun1,et al(1Dept of Infectious Diseases,The Second Affiliated Hospital of Anhui Medical University,Hefei 230601,2The Key Laboratory of Gene Resource Utilization for Severe Diseases,The Ministry of Education of Chinaand Anhui Province,The Key Laboratory of Zoonoses of Anhui Province,Hefei 230032) |
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| Abstract: | Objective To design and construct the artificial miRNA(amiRNA) vectors which targeted HBV,observe whether amiRNA-mediated RNA interference could decrease the replication level of HBV.Methods The amiRNA expression vectors targeted by HBV P and X gene were construted according to the pcDNA6.2-G/W-miR vector instruction.The plasmids were transient transfected by lipofectemine 2000 reagment.Semi-quantitative RT-PCR was utilized to observe suppression level of HBV mRNA;The MEIA was carried out to detect quantita... |
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| Keywords: | hepatitis virus B type RNA small interfering |
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