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Rolling-Circle Replication of an Animal Circovirus Genome in a Theta-Replicating Bacterial Plasmid in Escherichia coli

机译:在大肠杆菌中θ复制细菌质粒中的动物圆环病毒基因组的滚圆复制。

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摘要

A bacterial plasmid containing 1.75 copies of double-stranded porcine circovirus (PCV) DNA in tandem (0.8 copy of PCV type 1 [PCV1], 0.95 copy of PCV2) with two origins of DNA replication (Ori) yielded three different DNA species when transformed into Escherichia coli: the input construct, a unit-length chimeric PCV1Rep/PCV2Cap genome with a composite Ori but lacking the plasmid vector, and a molecule consisting of the remaining 0.75 copy PCV1Cap/PCV2Rep genome with a different composite Ori together with the bacterial plasmid. Replication of the input construct was presumably via the theta replication mechanism utilizing the ColE1 Ori, while characteristics of the other two DNA species, including a requirement of two PCV Oris and the virus-encoded replication initiator Rep protein, suggest they were generated via the rolling-circle copy-release mechanism. Interestingly, the PCV-encoded Rep′ protein essential for PCV DNA replication in mammalian cells was not required in bacteria. The fact that the Rep′ protein function(s) can be compensated by the bacterial replication machinery to support the PCV DNA replication process echoes previous suggestions that circular single-stranded DNA animal circoviruses, plant geminiviruses, and nanoviruses may have evolved from prokaryotic episomal replicons.
机译:包含1.75拷贝双链猪圆环病毒(PCV)DNA的细菌质粒(0.8拷贝PCV 1型[PCV1],0.95拷贝PCV2)串联,具有两个DNA复制起点(Ori),转化后可产生三种不同的DNA种类。进入大肠杆菌:输入构建体,具有复合Ori但缺少质粒载体的单位长度嵌合PCV1Rep / PCV2Cap基因组,以及一个分子,该分子由其余0.75个拷贝的PCV1Cap / PCV2Rep基因组与不同的复合Ori组成,并带有细菌质粒。输入构建体的复制大概是通过利用ColE1 Ori的theta复制机制进行的,而其他两个DNA物种的特征(包括需要两个PCV Oris和病毒编码的复制引发剂Rep蛋白)表明它们是通过滚动产生的圈复制释放机制。有趣的是,细菌中不需要PCV编码的Rep'蛋白,该蛋白对于在哺乳动物细胞中PCV DNA复制至关重要。 Rep'蛋白功能可以通过细菌复制机制补偿以支持PCV DNA复制过程这一事实与以前的建议相呼应,即环状单链DNA动物圆环病毒,植物双生病毒和纳米病毒可能是由原核附加型复制子进化而来的。

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  • 作者

    Cheung, Andrew K.;

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  • 年度 2006
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  • 原文格式 PDF
  • 正文语种 en
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