锤头核酶对鹅副粘病毒SF02在鸡胚成纤维细胞中复制的抑制作用

Jian Zou, Zun-Xun Gong
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引用次数: 0

摘要

设计了针对鹅副粘病毒SF02 (F)基因的锤头核酶RzF598及其功能失能突变体dRzF598。将核酶基因插入真核表达载体pcDNA3中,构建了pcDNA-RzF598和pcDNA-dRzF598转基因质粒。以缺乏完整核酶基因的质粒pcDNA3作为对照。将pcDNA-RzF598、pcDNA-dRzF598和pcDNA3质粒转染鸡胚成纤维细胞。检测感染CEFs释放的病毒浓度和CEFs的存活率。结果表明,RzF598成功抑制了SF02在CEFs中的复制。转染pcDNA-RzF598并感染SF02的CEFs的存活率高达78.8%,而未转染pcDNA3和转染SF02的CEFs感染SF02后的存活率仅为5%左右。
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Inhibition of replication of goose paramyxovirus SF02 by hammerhead ribozyme targeting to the SF02 F mRNA in chicken embryo fibroblasts.

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%.

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