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首页> 外文期刊>Journal of Virology >Anti-VPg antibody precipitation of product RNA synthesized in vitro by the poliovirus polymerase and host factor is mediated by VPg on the poliovirion RNA template.
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Anti-VPg antibody precipitation of product RNA synthesized in vitro by the poliovirus polymerase and host factor is mediated by VPg on the poliovirion RNA template.

机译：通过Poliovirus聚合酶和宿主因子在体外合成的产品RNA的抗VPG抗体沉淀由VPG介导PolioVirion RNA模板。

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摘要

Antibody to the poliovirus genome-linked protein, VPg, specifically immunoprecipitated the product RNA synthesized in vitro by the poliovirus RNA polymerase and HeLa cell host factor when VPg-linked poliovirion RNA was used as a template. The largest product RNA that was immunoprecipitated was twice the size of the template RNA. The complete denaturation of the product RNA with CH3HgOH had no effect on the immunoprecipitation reaction. In contrast, CH3HgOH denaturation prevented the immunoprecipitation of the oligo(U)-primed product RNA. Immunoprecipitation of the product RNA synthesized in the host-factor-dependent reaction was prevented if VPg was removed from the template RNA by pretreatment with proteinase K or if an RNA template without VPg was used in the reaction. The results support our previous evidence that a covalent linkage exists between the labeled negative-strand product RNA and the VPg-linked template RNA and suggest that the purified polymerase and host factor initiated RNA synthesis in vitro in the absence of VPg or a VPg-precursor protein.
机译：当VPG连接的脊髓尿RNA用作模板时，Poliovirus基因组连接蛋白，VPG，特别是免疫沉淀的脊髓灰质炎聚合酶和HeLa细胞宿主因子合成的产物RNA。免疫沉淀的最大产品RNA是模板RNA的尺寸的两倍。产物RNA与CH3HGoH的完全变性对免疫沉淀反应没有影响。相比之下，CH3HGoH变性阻止了寡核苷酸（U） - 预期产物RNA的免疫沉淀。防止在宿主因子依赖反应中合成的产物RNA，如果通过用蛋白酶K从模板RNA中除去VPG，或者如果在反应中使用没有VPG的RNA模板，则在模板RNA中除去VPG。结果支持我们以前的证据表明，标记的阴茎产物RNA和VPG连接的模板RNA之间存在共价连接，并表明纯化的聚合酶和宿主因子在不存在VPG或VPG-前体的情况下在体外引发RNA合成蛋白质。

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