RNA干扰对乙型肝炎病毒复制的体外影响
乙型肝炎病毒;RNA干扰;小发卡RNA;逆转录病毒载体;抗病毒治疗,王新宇,张继明,尹有宽,谢怡,黄玉仙,邬祥惠,翁心华,王新宇,通讯作者:,InhibitionofhepatitisBvirusexpressionandreplicationbyR
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王新宇, 张继明, 尹有宽, 谢怡, 黄玉仙, 邬祥惠, 翁心华, 复旦大学附属华山医院传染病科/华山医院肝病中心 上海市 200040
王新宇, 硕士, 医师, 主要从事病毒性肝炎的临床与基础研究.上海市科技攻关计划项目, No. 034119906
通讯作者: 张继明, 200040, 上海市乌鲁木齐中路12号, 复旦大学附属华山医院传染病科. jmzhang@fudan.edu.cn
电话: 021-62489999-6505 传真: 021-62486140
收稿日期: 2007-01-30 接受日期: 2007-02-13
Inhibition of hepatitis B virus expression and replication by RNA interference
Xin-Yu Wang, Ji-Ming Zhang, You-Kuan Yin, Yi Xie, Yu-Xian Huang, Xiang-Hui Wu, Xin-Hua Weng
Xin-Yu Wang, Ji-Ming Zhang, You-Kuan Yin, Yi Xie, Yu-Xian Huang, Xiang-Hui Wu, Xin-Hua Weng,Department of Infectious Diseases; Center for Liver Diseases, Huashan Hospital Affiliated to Fudan University, Shanghai 200040, China
Supported byShanghai Science and Technology Research and Development Program, No. 034119906
Correspondence to:Ji-Ming Zhang, Department of Infectious Diseases; Center for Liver Diseases, Huashan Hospital Affiliated to Fudan University, 12 Urumchi Middle Road, Shanghai 200040, China. jmzhang@fudan.edu.cn
Received:2007-01-30 Accepted:2007-02-13
Abstract
AIM: To study the RNA interference on hepatitis B virus (HBV) replication by a reverse transcription virus vector which can express short hairpin RNA inside cells.
METHODS: pSIREN vectors with inserted oligonucleotides targeting on reverse transcriptase (RT) regions of HBV genome were constructed. These plasmids were co-transfected with pHBV3.8 into Huh-7 cells. Viral antigens were measured by enzyme-linked immunosorbent assay (ELISA). HBV core particle DNA was measured and quantified by real-time fluorescence quantitative polymerase chain reaction (RFQ-PCR) and Southern blot. Viral RNA was analyzed by Northern blot.
RESULTS: Three RNA interfering targets were identified, and three corresponding retrovirus vectors, named 154i, 312i and 734i, were obtained. It was found that 312i markedly inhibited the expression of pHBV3 ......
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