用狂犬病毒核蛋白基因小干扰RNA库抑制狂犬病毒复制和感染的研究.pdfVIP

第 21 卷 3 期 中 国 病 毒 学 21 （3 ）：238-243 2006 年 5 月 VIROLOGICA SINICA May 2006 用狂犬病毒核蛋白基因小干扰 RNA 库抑制狂犬病毒 复制和感染的研究 王继麟，朱家鸿** ，徐葛林 （武汉生物制品研究所基因工程室，湖北武汉 430060 ） Inhibition of Rabies Virus Replication and Infection by Rabies Virus Nucleoprotein Gene siRNA Cocktail ** WANG Ji-lin ，ZHU Jia-hong ，XU Ge-lin (Wuhan Institute of Biological Products, Wuhan 430060, China) Abstract ：Small interference RNA mixtures (siRNA Cocktails ) of Rabies virus (RV) N gene, P gene and G gene were prepared by digesting long double-stranded RNA (dsRNA) with RNase Ⅲ. These siRNA cocktails were transfecteing into BSR or MNA cell monolayers and the direct immunofluores- cence assay (FA) was used to measure the replication and infection of RV on the cell monolayers. We found that the siRNA Cocktails of N gene can obviously and stably inhibit the replication and infection of RV. A descent of mRNA of N gene was also detected by RT-PCR. Under the same situation, the siRNA Cocktails of P gene or G gene showed no or weak inhibition to the replication and infection of RV. All of these results provide a basis for selecting target gene of RV in RNA itreatment and its further application. Key words ：Rabies virus (RV) ；siRNA Cocktail；Replication ；Inhibition 摘要：采用RNase Ⅲ消化长片段双链RNA 的方法，制备了狂犬病毒N 基因、P 基因和 G 基因小干扰RNA 库（siRNA Cocktail ）。将 siRNA 转染 BSR 和 MNA 细胞单层后，采用直接免疫荧光法观察发现，N 基因 siRNA Cocktail 能 够对 RV 的复制和感染产生明显和稳定的抑制作用，并且通过RT-PCR 在转录水平探测到 mRNA 产量的降低；而 P 基因或 G 基因 siR