期刊
NUCLEIC ACIDS RESEARCH
卷 42, 期 16, 页码 10596-10604出版社
OXFORD UNIV PRESS
DOI: 10.1093/nar/gku737
关键词
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资金
- David and Astrid Hagelen Foundation
- EU FP7 through EScoDNA ITN
- Vetenskapsradet [2013-5883]
- Stiftelsen for Strategisk Forskning (SSF) [FFL12-0219]
- Vetenskapsradet (Swedish Research Council)
- Swedish Foundation for Strategic Research (SSF) [FFL12-0219] Funding Source: Swedish Foundation for Strategic Research (SSF)
In rolling circle replication, a circular template of DNA is replicated as a long single-stranded DNA concatamer that spools off when a strand displacing polymerase traverses the circular template. The current view is that this type of replication can only produce single-stranded DNA, because the only 3'-ends available are the ones being replicated along the circular templates. In contrast to this view, we find that rolling circle replication in vitro generates large amounts of double stranded DNA and that the production of single-stranded DNA terminates after some time. These properties can be suppressed by adding single-stranded DNA-binding proteins to the reaction. We conclude that amodel in which the polymerase switches templates to the already produced single-stranded DNA, with an exponential distribution of template switching, can explain the observed data. From this, we also provide an estimate value of the switching rate constant.
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