Systematic Probing of the Sequence Selectivity of Exonuclease III with a Photosensitization Colorimetric Assay

Exonuclease III (Exo III) is an important enzymatic tool that is being widely used in molecular biology, biotechnology, and bioassay development. Exo III prefers to cleave double-stranded DNA (dsDNA) with blunt and recessed 3′-termini rather than their protruding counterpart. While it has been accepted that a short 3′-overhang (e.g., >4 nt) is necessary to protect a dsDNA from Exo III cleavage, critical roles of the length and sequence of this 3′-overhang remain unexplored. Herein, we develop a novel light-induced colorimetric assay allowing the systematic probe of the sequence selectivity of Exo III in a rapid and high-throughput manner. Our finding that Exo III is highly specific to 3′-overhang in terms of both length and sequence will be valuable for guiding the design of bioassays and DNA manipulating tools mediated by Exo III.

• Publication year

  2019

• Venue

  ACS Omega

• Publication date

  2019-08-06

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.1021/acsomega.9b01560PMID 31460466PMCID 6705212
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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