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线性质粒原噬菌体N15的repA复制基因的功能特性分析

Functional characterization of the repA replication gene of linear plasmid prophage N15.

作者信息

Mardanov Andrey V, Ravin Nikolai V

机构信息

Centre "Bioengineering", Russian Academy of Sciences, Prosp. 60-let Oktiabria, bld. 7-1, Moscow 117312, Russia.

出版信息

Res Microbiol. 2006 Mar;157(2):176-83. doi: 10.1016/j.resmic.2005.06.008. Epub 2005 Aug 8.

Abstract

The prophage of coliphage N15 is not integrated into the chromosome, but exists as a linear plasmid molecule with covalently closed ends. The only phage gene required for replication of circular N15 miniplasmids is repA (gene 37). Here we show that RepA-driven replication of the N15-based circular and linear miniplasmids is independent of host DnaB helicase protein, but requires the host DnaG primase. Replication of phage N15 DNA during lytic growth following infection does not depend on either DnaG or DnaB, but DnaG is required for lytic development after induction of the N15 lysogen. Finally, protein sequence analysis and replication data using different mutant strains suggest that RepA protein combines helicase and primase functions.

摘要

大肠杆菌噬菌体N15的原噬菌体不整合到染色体中,而是以具有共价封闭末端的线性质粒分子形式存在。环状N15小质粒复制所需的唯一噬菌体基因是repA(基因37)。我们在此表明,基于N15的环状和线性小质粒的RepA驱动复制独立于宿主DnaB解旋酶蛋白,但需要宿主DnaG引物酶。感染后裂解生长期间噬菌体N15 DNA的复制不依赖于DnaG或DnaB,但N15溶原菌诱导后的裂解发育需要DnaG。最后,使用不同突变菌株的蛋白质序列分析和复制数据表明,RepA蛋白兼具解旋酶和引物酶功能。

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