蛋白质起始的DNA病毒复制机制

Protein-Primed Mechanism of DNA Virus Replication

DNA聚合酶在DNA合成过程中需要的引物包括RNA引物、DNA自我引物和蛋白质引物3种类型。新DNA链(如冈崎片段)的复制多是在DNA模板上合成一段RNA引物,细小病毒利用其基因组末端的反向末端重复序列(ITRs)自我折叠成DNA引物,而一些DNA、RNA病毒及真菌质粒起始复制反应的引物则是蛋白质。以感染原核生物的噬菌体Phi29和真核DNA病毒腺病毒为例,从复制过程所涉及的蛋白质、对复制原点的识别、复制起始反应、新链的延伸、复制终止过程等方面详细阐述DNA病毒由蛋白质引发的复制机制,并对已商品化的Phi29 DNA聚合酶产品多重置换扩增及单细胞测序等的应用以及基于噬菌体Phi29蛋白质起始的最小复制系统体外扩增异源DNA等最新的应用研究作相关总结介绍。

DNA polymerase essential for the process of DNA synthesis comprises RNA primer, DNA self-primer, and protein primer three types of primers. The replication of new DNA strands （ e. g. Okazaki fragments） are mostly initia- ted with the synthesis of an RNA primer on the DNA template, Parvovirus using inverted terminal repeats （ ITRs ） at the end of its genome which self fold into a DNA primer, while some of the DNA, RNA viruses and fungal plasmids starting replication reaction with a protein primer. In this article, phage Phi29 and eukaryotic DNA virus adenovirus that infected prokaryote were taken as models and elaborate on their DNA virus replication mechanism primed by pro- tein, ranging from proteins involved in the replication process, recognition of the replication origin, initiation of repli- cation, DNA-primed elongation and termination of TP-DNA replication. The commercialized Phi29 DNA polymerase products applied for multiple displacement whole genome amplification （WGA） and single-cell sequencing, amplification of heterologous DNA with a minimal replication system based on phage Phi29 in vitro and other new applied researches were summarized and introduced reciprocally in this paper.