Baculovirus Replication: Purification and identification of the Trichoplusia ni Nuclear Polyhedrosis Virus-induced DNA Polymerase
Baculovirus Replication: Purification and identification of the Trichoplusia ni Nuclear Polyhedrosis Virus-induced DNA Polymerase
复制标题
杆状病毒复制：粉纹夜蛾核型多角体病毒诱导的 DNA 聚合酶的纯化和鉴定
DOI：
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发表时间：
1983
期刊：
影响因子：
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通讯作者：
D. Kelly
中科院分区：
文献类型：
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作者：
X. Wang;D. Kelly
总结
用DNA-纤维素和磷酸纤维素柱层析法分析了粉纹夜蛾核型多角体病毒（Trichoplusia ni multiple nucleocapsidNuclear polyhedrosis virus，NDPV）感染草地贪夜蛾（Spodoptera frugiperda）细胞的DNA聚合酶活性。在受感染的细胞中，发现一种新的活性组分与柱的结合比未感染细胞中的聚合酶活性更强。通过粗提物的DNA-纤维素层析和半纯化活性的磷酸-纤维素层析的组合纯化感染细胞特异性DNA聚合酶。最终产物含有分子量为126 000的单一多肽，这在未感染的细胞中没有发现。纯化的酶能被阿非迪霉素和[E]-5-（2-溴乙烯基）-2′-脱氧尿苷三磷酸抑制，而不被溴乙烯基脱氧尿苷抑制。该酶被证明是一种早期酶，可能是一种延迟的早期蛋白质，因为它存在于被阿非迪霉素抑制的细胞中，这些细胞被药物锁定在早期蛋白质的合成中。
Summary
DNA polymerase activity present in Trichoplusia ni multiple nucleocapsid nuclear polyhedrosis virus-infected Spodoptera frugiperda cells has been analysed by chromatography on DNA-cellulose and phosphocellulose columns. In infected cells a new fraction of activity was found to bind to the columns more strongly that did polymerase activity in uninfected cells. The infected-cell-specific DNA polymerase was purified by a combination of DNA-cellulose chromatography of crude extracts and phospho-cellulose chromatography of the semi-purified activity. The final product contained a single polypeptide of molecular weight 126 000 which was not found in uninfected cells. The purified enzyme was inhibited by aphidicolin and [E]-5-(2-bromovinyl)-2′-deoxyuridine triphosphate, but not by bromovinyldeoxyuridine. The enzyme was shown to be an early enzyme, probably a delayed early protein, since it was present in cells inhibited by aphidicolin which were locked into the synthesis of early proteins by the drug.