RNA干扰用于柯萨奇病毒B3感染治疗作用研究  

Effects of RNAi against CVB3 infection

作　　者：陈洁[1] 何嘉莉[1] 陈佳琪[2] 戴玲珊[1] 陈佳玉[1] 

机构地区：[1]台州学院医学院,浙江台州318000 [2]台州市立医院,浙江台州318000

出　　处：《中国热带医学》2013年第11期1303-1305,共3页China Tropical Medicine

基　　金：国家级大学生创新训练计划项目(No.201310350015);浙江省自然科学基金(No.Y2100248);浙江省科技厅项目(No.2009C33155);浙江省卫生厅项目(No.2009A218);台州市科技局项目(No.102KY15);浙江省中医药管理局项目(No.2011ZA113)

摘　　要：目的研究观察自行构建的siRNA表达载体对柯萨奇病毒B3(CVB3)感染的潜在治疗作用。方法以CVB3聚合酶编码区基因序列为靶点,构建发夹型siRNA真核表达载体pGCsi-U6/GFP/P及与CVB基因组序列无关的阴性对照pGCsi-U6/GFP/N。利用脂质体转染进入Hela细胞后,用CVB3毒株进行攻击,观察感染后Hela细胞病变情况,并用RT-PCR法检测细胞培养上清中的CVB3负链RNA水平。结果构建的表达质粒pGCsi-U6/GFP/P在Hela细胞能够正确表达,其表达产物能够使CVB3基因沉默,并间接抑制了CVB3的复制。结论本实验成功构建了能够抑制CVB3的发夹状siRNA表达质粒,该质粒具有增强细胞抗CVB3感染的能力。Objective To investigate the anti-CVB3 effect of the siRNA expressing vector which we constructed. Methods Hairpin siRNA expression vector pGCsi-U6/GFP/P and pGCsi-U6/GFP/N （Negative control has no relationship with CVB3 gene sequence） were constructed according to the polymerase gene.The ceils were attacked with CVB3 after transfected them into Hela cells, and the condition of these cells were observed. The negative strand RNA of CVB3 in the cell culture supernatant determined by RT-PCR and the therapeutic effect of the product produced was analyzed by the siRNA expression vector. Results The expressional plasmid pGCsi-U6/GFP/Pwas constructed and this plasmid could express correctly in Hela cells and it could induce CVB3 silence and inhibit the replicatian of CVB3 effectively. Conclusions The CVB3 hairpin siRNA expressional plasmid was successfully constructed,and this plasmid could enhance the capabihty of cells against CVB3 infection. The success of this experiment has laid a solid foundation for treatment of CVB3 infection and the study of proteins functional.

关 键 词：柯萨奇病毒B3 HELA细胞 RNA干扰 

分 类 号：R373.23[医药卫生—病原生物学]