Difference between revisions of "Part:BBa K4674002"

 
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<partinfo>BBa_K4674002 short</partinfo>
 
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The RepA is a replication initiator protein that initiate the process of rolling circle replication (RCR) of the R-plasmid, pC194. RepA binds to the double-stranded origin (DSO) and creats nick on one of the DNA strand. The DNA polymerase use unnicked strand as template to elongate the nicked strand. Finally, the elongated nicked strand is completely replaced by the newly synthesised strand, and the replication initiator enzyme will then ligate the elongated nicked strand into a circular DNA.
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<strong> The RCR mechanism: RepA protein, RCORI105 and RCORI65 </strong>
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Rolling Circle Replication (RCR) is a fundamental molecular process that plays an important role in DNA replication and various molecular biology applications. It involves one-way amplification of circular DNA samples, producing long single-stranded or double-stranded DNA products with repetitive sequences.
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In RCR, each replication initiator enzyme (e.g. RepA) recognizes its corresponding double-stranded origin (DSO) of the target plasmid (e.g. pC194), and creates a nick on one of the DNA strands. The DNA polymerase uses an unnicked strand as a template to elongate the nicked strand. Finally, the elongated nicked strand is completely replaced by the newly synthesised strand, and the replication initiator enzyme will then ligate the elongated nicked strand into a circular ssDNA. The circular ssDNA could server as a template for secondard strand synthesis (Ruiz-Masó et al 2015).
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https://static.igem.wiki/teams/4674/wiki/1-rolling-circle-replication-2.png
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Fig. The mechanism of RCR replication
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<strong> Experimental result </strong>
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Revision as of 06:23, 12 October 2023


RepA

The RCR mechanism: RepA protein, RCORI105 and RCORI65

Rolling Circle Replication (RCR) is a fundamental molecular process that plays an important role in DNA replication and various molecular biology applications. It involves one-way amplification of circular DNA samples, producing long single-stranded or double-stranded DNA products with repetitive sequences.

In RCR, each replication initiator enzyme (e.g. RepA) recognizes its corresponding double-stranded origin (DSO) of the target plasmid (e.g. pC194), and creates a nick on one of the DNA strands. The DNA polymerase uses an unnicked strand as a template to elongate the nicked strand. Finally, the elongated nicked strand is completely replaced by the newly synthesised strand, and the replication initiator enzyme will then ligate the elongated nicked strand into a circular ssDNA. The circular ssDNA could server as a template for secondard strand synthesis (Ruiz-Masó et al 2015).

1-rolling-circle-replication-2.png

Fig. The mechanism of RCR replication

Experimental result



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]