gp64基因相应dsRNA对家蚕核型多角体病毒(BmNPV)增殖的抑制  

Inhibitory Effects of the Corresponding dsRNA of gp64 Gene on BmNPV Multiplication

作　　者：夏定国[1] 张国政[1] 王文兵[2] 赵巧玲[1] 唐顺明[1] 

机构地区：[1]中国农业科学院蚕业研究所/农业部家蚕生物技术重点开放实验室 [2]江苏大学生命科学研究院,镇江212013

出　　处：《中国农业科学》2007年第12期2882-2887,共6页Scientia Agricultura Sinica

基　　金：江苏省自然科学基金前期预研重大项目(BK2004206)

摘　　要：【目的】研究gp64基因相对应的多个dsRNA对家蚕核型多角体病毒增殖的抑制效果。【方法】体外合成dsRNA,通过病毒滴度试验、实时定量RT-PCR法研究gp64基因的不同区域、不同长度与RNAi干扰效果的关系。【结果】供试的6个dsRNA的最大抑制效果相当(滴度TCID50的最大抑制差值为4.00左右);经RNAi的不同时间点的gp64mRNA表达水平均明显下调(P<0.01),其中48h的gp64mRNA的表达量约为对照的1/300。【结论】6个dsRNA均能有效地抑制病毒的基因表达和增殖;gp64ORF第1390～1499位点(G3-3)是RNAi的较佳靶位点。【Objective】Studies were performed to investigate the inhibitory effects of different corresponding dsRNAs of gene gp64 on the multiplication of Bombyx mori Nucleopolyhedrovirus (BmNPV). 【Method】 DsRNA were synthesized in vitro .Using the methods of measuring virus titer and real-time fluorescent quantitative RT-PCR, the interference effect of different target domain or length of gp64 gene were tested.【Result】The maximum inhibitory effect of the six dsRNAs fragments showed no obvious difference (the maximum distinction of virus titer TCID50 was about 4.00). At different periods of RNA interference, the expression level of gp64 mRNA were down regulated (P<0.01). At 48 hpt, the expression amount of gp64 mRNA was about 1/300 of the control .【Conclusion】These six dsRNAs can inhibit the expression of viral gene and the multiplication of BmNPV. The best target site for RNAi is located at 1 390-1 499 in gp64 ORF.

关 键 词：双链RNA 家蚕核型多角体病毒 gp64基因 病毒滴度 实时定量RT-PCR 

分 类 号：S884.51[农业科学—特种经济动物饲养]