In vitro synthesis of bacteriophage phi X174 by purified components.
In vitro synthesis of bacteriophage phi X174 by purified components.
复制标题
通过纯化组分体外合成噬菌体 phi X174。
DOI：
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发表时间：
1983
影响因子：
11.1
通讯作者：
M. Hayashi
中科院分区：
文献类型：
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作者：
A. Aoyama;R. Hamatake;M. Hayashi
从纯化的组分重构能够合成感染性phi X174噬菌体颗粒的体外系统。该合成需要phi X174超螺旋复制形式DNA、phi X174编码的蛋白质A、C、J和prohead、大肠杆菌DNA聚合酶III全酶、rep蛋白和脱氧尿苷三磷酸酶（dUTPase，dUTP核苷酸水解酶，EC 3.6.1.23）以及MgCl2、四种脱氧核苷三磷酸和ATP。噬菌体生产与病毒单链DNA的合成偶联。超过70%的合成颗粒在蔗糖梯度中沉淀在成熟噬菌体的位置，并与感染性相关。对宿主蛋白质的简单要求表明，噬菌体合成反应中病毒链合成的机制类似于复制型DNA复制过程中病毒链合成的机制。
An in vitro system capable of synthesizing infectious phi X174 phage particles was reconstituted from purified components. The synthesis required phi X174 supercoiled replicative form DNA, phi X174-encoded proteins A, C, J, and prohead, Escherichia coli DNA polymerase III holoenzyme, rep protein, and deoxyuridinetriphosphatase (dUTPase, dUTP nucleotidohydrolase, EC 3.6.1.23) as well as MgCl2, four deoxyribonucleoside triphosphates, and ATP. Phage production was coupled to the synthesis of viral single-stranded DNA. More than 70% of the synthesized particles sedimented at the position of mature phage in a sucrose gradient and associated with the infectivity. The simple requirement of the host proteins suggests that the mechanism of viral strand synthesis in the phage-synthesizing reaction resembles that of viral strand synthesis during the replication of replicative form DNA.