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

ABSTRACT: 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.

SUBMITTER: Ding Y

PROVIDER: S-EPMC6705212 | biostudies-literature | 2019 Aug

REPOSITORIES: biostudies-literature

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

Ding Yu Y Li Xianming X Zhang Xinfeng X Li Feng F Hou Xiandeng X Wu Peng P

ACS omega 20190806 8

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 ...[more]

PMID: 31460466

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