Oligonucleotide-Directed Nucleic Acid Scission by Micrococcal Nuclease

Ralf Landgraf, Chi hong B. Chen, David S. Sigman

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

"Sequence-dictated" scission of DNA can be achieved by tethering a fusion protein composed of glutathione S-transferase and attenuated micrococcal nuclease (MN) to a targeting oligonucleotide using Cibacron blue (CB) F3G-A. Deoxyoligonucleotides derivatized with this dye bind to the fusion protein in gel mobility shift assays. This binding scheme was successfully used to achieve site-specific scission of a single-stranded DNA substrate after hybridization with a CB-derivatized complementary oligonucleotide. Although covalently cross-linked hybrids of MN and oligonucleotides have been successfully used in the past to target nucleolytic activity, this novel scheme opens new possibilities for targeting and probing both DNA and RNA sequences by allowing the addition of the nuclease subsequent to hybridization.

Original languageEnglish (US)
Pages (from-to)10607-10615
Number of pages9
JournalBiochemistry
Volume33
Issue number35
DOIs
StatePublished - Sep 1 1994
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry

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