Isolation and subunit structure of polycytidylate-dependent RNA polymerase of encephalomyocarditis virus

脑心肌炎病毒多胞苷酸依赖性RNA聚合酶的分离和亚基结构

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Abstract

A polycytidylate-dependent RNA polymerase of encephalomyocarditis virus was isolated from infected BHK 21 cells. The enzyme was associated with a smooth-membrane fraction, from which it was extracted by a mixture of sodium dodecyl sulfate, Triton X-100, and dithiothreitol, and further purified by chromatography on a Dowex-1 column and by glycerol gradient sedimentation. Analysis of a 6S glycerol gradient peak of RNA polymerase activity by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed the presence of five polypeptides, of molecular weights 72,000, 65,000, 57,000, 45,000, and 35,000. The molecular weights of four of the polypeptides (72,000, 65,000, 45,000, and 35,000) are almost identical to the reported molecular weights of the four subunits of Qbeta replicase.

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