IN VITRO TRANSLATION AS A PROBE FOR THE MECHANISM OF POLIOVIRUS ATTENUATION

体外翻译作为脊髓灰质炎病毒减毒机制的探针

• 批准号: 3748141
• 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/3748141
• 关键词: HeLa cells; attenuated microorganism; chemical stability; genetic strain; genetic translation; genome; nucleic acid sequence; point mutation; poliomyelitis vaccine; poliovirus; virion; virulence; virus RNA; virus genetics; virus replication

Attenuation of poliovirus neurovirulence is the result of a limited number of nucleotide changes in the poliovirus genome. A major site for attenuation of Type 3 poliovirus (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. Mola, Paul, and Wimmer,Science, 254, 1647-1651(1991) have reported that an in vitro translation system derived from uninfected HeLa cells translated added poliovirus RNA in a manner that closely mimics the full viral replicative cycle in human cells. More recently, Barton and Flanegan, J. of Virology, 67, 822- 831, (1993), have provided additional information about this system. We are using this system to determine whether the genomic changes that decrease neurovirulence exhibit an effect on the in vitro translation of virion RNA. The response of HeLa cell extracts to the addition of wild-type poliovirus RNA are being evaluated and will be used to compare the response to RNA derived from the attenuated Sabin strain. The goal of this project is to obtain further information concerning the function of the nucleotide changes that result in attenuation of the poliovirus genome. Understanding the interaction of these sequences with viral and host factors could lead to further changes in the attenuated genome that would result in increased stabilty of the attenuated phenotype.

脊髓灰质炎病毒神经毒力的减弱是由于 脊髓灰质炎病毒基因组中核苷酸变化的数量。 一个主要的网站， 3型脊髓灰质炎病毒（472位）的减毒发生在非编码 基因组中与病毒调节有关的区域 翻译. 翻译中的变化是如何（以及是否）导致 脊髓灰质炎病毒衰减仍有待确定。 莫拉，保罗， 维默（Wimmer），科学（Science），254，1647-1651（1991 来源于未感染的HeLa细胞的翻译系统 脊髓灰质炎病毒RNA的方式，密切模仿完整的病毒复制 人体细胞的周期 最近，巴顿和Flanegan，J. Virology，67，822- 831，（1993）提供了额外的信息 关于这个系统。 我们正在使用这个系统来确定 降低神经毒力的基因组变化对神经系统的影响 病毒体RNA的体外翻译。 HeLa细胞提取物对 目前正在对添加野生型脊髓灰质炎病毒RNA进行评估， 用于比较对来自减毒Sabin的RNA的反应 株 这个项目的目标是获得更多的信息 关于核苷酸改变的功能， 脊髓灰质炎病毒基因组的减毒。 理解的相互作用 这些带有病毒和宿主因子的序列可能会导致进一步的变化 在减毒的基因组中，这将导致 减毒表型