IN VITRO TRANSLATION OF WILD-TYPE AND ATTENUATED POLIOVIRUS GENOMES

脊髓灰质炎病毒神经毒力的减弱是有限数量的 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节 如何（以及如果） 翻译的变化导致脊髓灰质炎病毒减毒仍然是 确立了习 Mola、Paul和维默的最新报告，《科学》1991;254：1647-1651 表明源自HeLa细胞的体外翻译系统 翻译添加的脊髓灰质炎病毒RNA的方式，密切模仿完整的 病毒在人体细胞中的复制周期。 我们用这个系统 确定野生型和减毒基因组是否表现出不同的 在来自人细胞系的体外提取物中的翻译应答。 已经制备了HeLa细胞提取物， 通过体外合成脊髓灰质炎病毒蛋白质来合成脊髓灰质炎病毒RNA。 正在评价对野生型病毒RNA的应答特征 并将用于比较对携带减毒RNA的应答。 表型。 阐明翻译在脊髓灰质炎病毒减毒中的作用 神经毒力可能会导致进一步改善稳定性， 衰减表型。

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.