Emerson S U, Wagner R R
J Virol. 1973 Dec;12(6):1325-35. doi: 10.1128/JVI.12.6.1325-1335.1973.
The endogenous transcriptase present in purified vesicular stomatitis (VS) virions was solubilized with a Triton X-100 high-salt solution. The polymerase activity was purified on glycerol gradients and by phosphocellulose column chromatography; the viral proteins present in the active enzyme fractions were identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis. It was demonstrated that L protein, but not NS protein, was required for in vitro RNA synthesis on the VS viral nucleocapsid template. Solubilized L protein rebinds to the ribonucleoprotein template when the transcription complex is reconstituted, and the RNA synthesized in vitro by purified L protein hybridizes to virion RNA. Cyanogen bromide peptide fingerprints indicate that the large L protein is a unique polypeptide chain. It is concluded that the L protein functions as the transcriptase, and the nucleocapsid NS protein is not essential for in vitro RNA synthesis.
用Triton X-100高盐溶液溶解纯化的水泡性口炎（VS）病毒粒子中存在的内源性转录酶。通过甘油梯度和磷酸纤维素柱色谱法纯化聚合酶活性；通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳鉴定活性酶组分中存在的病毒蛋白。结果表明，在VS病毒核衣壳模板上进行体外RNA合成需要L蛋白，而不是NS蛋白。当转录复合物重组时，溶解的L蛋白重新结合到核糖核蛋白模板上，并且纯化的L蛋白在体外合成的RNA与病毒粒子RNA杂交。溴化氰肽指纹图谱表明大L蛋白是一条独特的多肽链。得出的结论是，L蛋白作为转录酶起作用，核衣壳NS蛋白对于体外RNA合成不是必需的。
