Selection of genetic inhibitors of rabies virus
Selection of genetic inhibitors of rabies virus
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DOI：
10.1007/s00705-004-0299-6
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发表时间：
2004-08-01
影响因子：
2.7
通讯作者：
Curtis, PJ
中科院分区：
文献类型：
--
作者：
Wunner, WH;Pallatroni, C;Curtis, PJ
来自狂犬病病毒基因组的五个基因中的四个基因的短随机片段的cDNA文库已被用于分离细胞内表达的抑制狂犬病病毒复制的遗传抑制因子（GSE）。两个核苷酸片段，一个来自狂犬病病毒核衣壳蛋白（N）基因，另一个来自磷蛋白（P）基因，已被确定为狂犬病病毒在细胞培养物中复制的抑制剂。N基因cDNA片段以有义方向表达，可产生影响病毒复制的显性负性蛋白。P cDNA片段以抑制性反义方向表达。使用ELISA检测细胞表面狂犬病病毒糖蛋白表达和免疫荧光检测细胞内狂犬病病毒N表达，检测细胞培养物中狂犬病病毒复制的抑制。正义和反义GSE由于其靶向抑制狂犬病病毒复制，在合理设计用于治疗狂犬病的抗病毒化合物中具有可能的用途。这种方法可以应用于任何病毒，特别是那些裂解其靶宿主细胞的病毒。
A cDNA library of short random fragments derived from four of the five genes of the rabies virus genome has been used to isolate genetic suppressor elements (GSEs) expressed intracellularly that inhibit rabies virus replication. Two nucleotide fragments, one from the rabies virus nucleocapsid protein (N) gene and the other from the phosphoprotein (P) gene, have been identified as inhibitors of rabies virus replication in cell culture. The N cDNA fragment is expressed in sense-orientation and could produce a dominant negative protein affecting virus replication. The P cDNA fragment is expressed in the inhibitory antisense direction. Inhibition of rabies virus replication was detected in cell culture using an ELISA for detection of rabies virus glycoprotein expression on the cell surface and immunofluorescence for detection of intracellular rabies virus N expression. Both the sense and antisense GSEs, because of their targeted inhibition of rabies virus replication, have possible uses in rational design of antiviral compounds for treatment of rabies. This approach could be applied to any virus, particularly to those that lyse their target host cell.