锤头状核酶在鸡胚成纤维细胞中的对鹅副粘病毒SF02繁殖的抑制作用(英文)  

Inhibition of Replication of Goose Paramyxovirus SF02 by Hammerhead Ribozyme Targeting to the SF02 F mRNA in Chicken Embryo Fibroblasts

作　　者：邹键[1] 龚祖埙[1] 

机构地区：[1]中国科学院上海生命科学研究院生物化学与细胞生物学研究所蛋白质组学重点实验室,上海200031

出　　处：《生物化学与生物物理学报》2003年第9期801-805,共5页

基　　金：上海市科技兴农重点攻关项目~~

摘　　要：针对鹅副粘病毒SF0 2的F基因 ,设计了锤头状核酶RzF5 98,并插入真核细胞表达载体pcDNA3获得该核酶的转基因质粒pcDNA RzF5 98。以无核酶功能的突变体dRzF5 98和pcDNA3空质粒为对照 ,分析核酶对病毒繁殖的抑制作用。结果表明 ,RzF5 98不仅能在体外成功切割靶基因的mRNA ,还能有效抑制SF0 2在鸡胚成纤维细胞中的繁殖。转染pcDNA RzF5 98的鸡胚成纤维细胞经SF0 2感染后存活率可达 80 %左右 ,而未转染任何质粒的细胞和转染pcDNA3空质粒的细胞经SF0 2感染后存活率只有约 5 %。转染dRzF5 98的细胞对病毒也具有一定抗性 ,这可能是由于反义RNA的作用。Hammerhead ribozyme RzF598 and its dysfunctional mutant dRzF598 targeting to the F gene of goose paramyxovirus SF02 have been designed. The transgenic plasmids pcDNA-RzF598 and pcDNA-dRzF598 were constructed by inserting ribozyme genes into eukaryotic expression vector pcDNA3. The plasmid pcDNA3 that lacks full ribozyme gene was used as a control. Plasmids pcDNA-RzF598, pcDNA-dRzF598 and pcDNA3 were transfected into chicken embryo fibroblasts (CEFs). The concentration of virus released by infected CEFs and the survival percentages of CEFs were identified. The results indicated that RzF598 successfully suppressed the replication of SF02 in CEFs. Survival percentage of CEFs being transfected with pcDNA-RzF598 and infected SF02 was up to 78.8%, while the survival percentages of untransfected CEFs and CEFs transfected with pcDNA3 after infection with SF02 were only about 5%.

关 键 词：锤头状核酶 鸡胚成纤维细胞 鹅副粘病毒 繁殖 新城疫病毒 

分 类 号：S852.2[农业科学—基础兽医学]