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As a summary, a typical DNA rolling circle replication has five steps: [2] Circular dsDNA will be "nicked". The 3' end is elongated using "unnicked" DNA as leading strand (template); 5' end is displaced. Displaced DNA is a lagging strand and is made double stranded via a series of Okazaki fragments. Replication of both "unnicked" and displaced ...
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).
Viral replication is nuclear. Entry into the host cell is achieved by penetration into the host cell. Replication follows the ssDNA rolling circle model. DNA templated transcription, with some alternative splicing mechanism is the method of transcription. The virus exits the host cell by nuclear egress, and nuclear pore export.
The virus replicates through an dsDNA intermediate initiated by the Rep protein. Two major genes are transcribed from open reading frame (ORF) 1 and 2. ORF1 encodes Rep and Rep' for initiation of rolling-circle replication; ORF2 encodes Cap, the only structural and most immunogenic protein forming the viral capsid. [15]
DNA nanoballs are simply formed by denaturing double stranded, adapter ligated libraries and ligating the forward strand only to a splint oligonucleotide to form a ssDNA circle. Faithful copies of the circles containing the DNA insert are produced utilizing Rolling Circle Amplification that generates approximately 300–500 copies.
[1] [2] [3] Diseases associated with this family include: bright yellow mosaic, yellow mosaic, yellow mottle, leaf curling, stunting, streaks, reduced yields. [ 2 ] [ 4 ] They have single-stranded circular DNA genomes encoding genes that diverge in both directions from a virion strand origin of replication (i.e. geminivirus genomes are ambisense ).
For imaging systems which cannot detect single fluorescence events, amplification of DNA templates is required. The three most common amplification methods are emulsion PCR (emPCR), rolling circle and solid-phase amplification. The final distribution of templates can be spatially random or on a grid.
A concatemer is a long continuous DNA molecule that contains multiple copies of the same DNA sequence linked in series. These polymeric molecules are usually copies of an entire genome linked end to end and separated by cos sites (a protein binding nucleotide sequence that occurs once in each copy of the genome).