特异性干扰PB-1基因对H5N1高致病性禽流感病毒复制的影响  

Inhibition of Highly Pathogenic Avian Influenza Virus of H5N1 Subtype by the siRNA Specific for PB-1 Gene

作　　者：张伟[1] 王铁成[2] 王承宇[2] 杨松涛[2] 高玉伟[2] 夏咸柱[2] 

机构地区：[1]大连医科大学微生物学教研室,中国辽宁大连116044 [2]军事医学科学院军事兽医研究所,中国吉林长春130062

出　　处：《生命科学研究》2013年第3期238-242,共5页Life Science Research

基　　金：国家科技支撑计划项目(2006BAD06A14)

摘　　要：针对H5N1高致病性禽流感病毒PB-1基因,通过RNA干扰技术,设计siRNA,研究其抑制病毒复制的效果.siRNA转染MDCK细胞并接毒后,病毒滴度测定、Real-time PCR和间接免疫荧光试验结果显示,所设计的siRNA(ps-PB1-777)具有明显的抑制病毒复制的效果,PB-1基因的拷贝数和病毒蛋白表达都下降.设计的siRNA可显著抑制高致病性禽流感病毒的复制.因此,本研究中PB-1基因的有效siRNA可为预防和治疗H5N1高致病性禽流感提供合理的技术支持和物质储备.PBI-specific siRNAs of HSN1 highly pathogenic avian influenza virus （HPAIV） were designed and detected for their abilities to inhibit virus replication in vitro. Two siRNAs （ps-PB1-777 and ps-PB1- 1282） were designed and constructed expression plasmids. Virus titer assay, Real-time PCR and indirect im- muno-fluorescence experiment were used to observe their inhibition effect in MDCK cells challenged with HSN1 HPAIV. The results suggested virus titers of ps-PB1-777 were lower （P〈0.05, 8 times comparing with the control group）; after treatment of ps-PB1-777, the levels of PB-1 mRNA greatly decreased （P〈0.05）; the levels of proteins of HSN1 HPAIV decreased. However, ps-PBl-1282 did not show the abilities to antiviral effect. As a result, ps-PB1-777 can effectively suppressed the replication of HSN1 HPAIV, and the findings might have significant applications for prophylaxis and therapy of influenza virus.

关 键 词：高致病性禽流感病毒(HPAIV) 禽流感 干扰RNA PB-1基因 

分 类 号：R373.13[医药卫生—病原生物学]