【摘要】 采用RNaseⅢ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(siRNACocktail)。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siRNACocktail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siRNACocktail对RV的复制和感染不产生或仅产生弱的抑制作用。这一结果为RNA干扰在RV研究中靶基因的选择及进一步的应用提供了依据。更多还原

【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.更多还原