IN VITRO TRANSLATION OF WILD-TYPE AND ATTENUATED POLIOVIRUS GENOMES

野生型和减毒型脊髓灰质炎病毒基因组的体外翻译

• 批准号: 3792548
• 负责人: R E LUNDQUIST
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3792548
• 关键词: HeLa cells; attenuated microorganism; genetic translation; genome; nucleic acid sequence; poliomyelitis vaccine; poliovirus; protein biosynthesis; tissue /cell culture; virulence; virus RNA; virus genetics; virus protein

Attenuation of poliovirus neurovirulence is the result of a limited number of nucleotide changes in the poliovirus genome. A major site for attenuation (position 472) occurs in a noncoding region of the genome that is involved in the regulation of viral translation. Exactly how (and if) changes in translation result in poliovirus attenuation remain to be established. A recent report by Mola, Paul, and Wimmer, Science 1991;254:1647-1651 suggested that an in vitro translation system derived from HeLa cells translated added poliovirus RNA in a manner that closely mimics the full viral replicative cycle in human cells. We are using this system to determine if wild-type and attenuated genomes exhibit different translational responses in the in vitro extract from a human cell line. HeLa cell extracts have been prepared which respond to the addition of poliovirus RNA by synthesizing poliovirus proteins in vitro. Characteristics of the response to wild type viral RNA are being evaluated and will be used to compare the response to RNA carrying the attenuated phenotype. Elucidation of the role translation plays in the attenuation of poliovirus neurovirulence could lead to further improvements in the stabilization of the attenuation phenotype.

脊髓灰质炎病毒神经毒力的减弱是有限数量的结果 脊髓灰质炎病毒基因组的核苷酸变化。一个主要的网站 衰减(位置472)发生在基因组的非编码区 参与了病毒翻译的调控。具体是如何(以及是否) 翻译的变化导致脊髓灰质炎病毒的减弱仍有待于 已经成立了。 Mola、Paul和Wimmer最近的一份报告，《科学》1991；254：1647-1651 提示从HeLa细胞衍生的体外翻译系统 翻译后的添加脊髓灰质炎病毒RNA的方式与完整的 人类细胞中的病毒复制周期。我们正在使用这个系统来 确定野生型和弱毒基因组是否表现出不同 人类细胞系体外提取液中的翻译反应。 已经制备了HeLa细胞提取物，它对添加 通过在体外合成脊髓灰质炎病毒蛋白来获得脊髓灰质炎病毒RNA。 对野生型病毒rna的反应特征正在评估中。 并将用于比较携带衰减的RNA的反应 表型。 阐明翻译在脊髓灰质炎病毒减毒中的作用 神经毒力可能导致进一步改善病毒的稳定性 衰减表型。