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What is the sequence which identifies an origin of replication and please detail an experiment that...

What is the sequence which identifies an origin of replication and please detail an experiment that determined the actual nucleotide sequence, from beginning to end.

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The site of origin of replication contains A-T-rich sequence that is a DNA unwinding element, and DnaA recognition sequence TGTGGATAA and 5’ TTATC/ACACA 3’ as the initiator sequence.

In eukaryotes, DNA replication origins contain autonomous replication sequences filled with autonomous consensus sequences. This consensus sequence is 5’/(A/T)TTTA(T/C)(A/G)TTT(A/T) 3’. GC rich sequence is also another sequence for the origin of replication.

To identify the sequence, use the following procedure:

  • Isolate DNA from a sample.
  • Perform restriction digestion of the sample.
  • Run the sample on the gel electrophoresis.
  • Perform southern blotting to identify the OriC sequence using the specific DNA probes.
  • Select out the particular DNA sequence bound to the antibody.
  • Sequence the selected DNA using Single molecule real time (SMRT) sequencing, or Nanopore DNA sequencing.

gladiator answered 2 years ago
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