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Rolling circle replication (RCR) is a process of unidirectional nucleic acid replication that can rapidly synthesize multiple copies of circular molecules of DNA or RNA, such as plasmids, the genomes of bacteriophages, and the circular RNA genome of viroids. Some eukaryotic viruses also replicate their DNA or RNA via the rolling circle mechanism.
This is accomplished by rolling circle replication with the Phi 29 DNA polymerase which binds and replicates the DNA template. The newly synthesized strand is released from the circular template, resulting in a long single-stranded DNA comprising several head-to-tail copies of the circular template. [ 10 ]
In the next step, the DNA molecules are circularized with T-tailed 30 bp long synthetic oligonucleotides (T30), which contains two outward-facing MmeI recognition sites, and the resulting circularized DNA undergoes rolling circle replication. The amplified circularized DNA molecules are then digested with MmeI (type IIs restriction endonuclease ...
In the single stranded DNA viruses—a group that includes the circoviruses, the geminiviruses, the parvoviruses and others—and also the many phages and plasmids that use the rolling circle replication (RCR) mechanism, the RCR endonuclease creates a nick in the genome strand (single stranded viruses) or one of the DNA strands (plasmids).
In its natural host (S. cerevisiae) the Flp/FRT system enables replication of a "2μ plasmid" by the inversion of a segment that is flanked by two identical, but oppositely oriented FRT sites ("flippase" activity). This inversion changes the relative orientation of replication forks within the plasmid enabling "rolling circle"—amplification ...
Rolling circle replication of a circular DNA plasmid. Helitrons are also a group of eukaryotic class II TEs. Helitrons do not follow the classical "cut and paste" mechanism. Instead, they are hypothesized to move around the genome via a rolling circle like mechanism.
Viral HUH endonucleases are involved in initiating rolling circle replication while ones of bacterial origin initiate bacterial conjugation. In biotechnology, they can be used to create protein-DNA linkages, [2] akin to other methods such as SNAP-tag. In doing so, they create a 5' covalent bond between the ssDNA and the protein.
The self-cleavage reactions, first reported in 1986, [5] [6] are part of a rolling circle replication mechanism. The hammerhead sequence is sufficient for self-cleavage [7] and acts by forming a conserved three-dimensional tertiary structure.