Cleavage of single strand oligonucleotides and bacteriophage phi X174 DNA by Msp I endonuclease

J Biol Chem. 1980 Nov 25;255(22):10559-62.

Abstract

Type II restriction endonucleases cleave duplex DNA at nucleotide sequences displaying 2-fold symmetry. Our data show that Msp I cleaves single strand oligonucleotides, d(G-A-A-C-C-G-G-A-G-A) and d(T-C-T-C-C-G-G-T-T) at 4 degrees, 25 degrees, and 37 degrees C reaction temperatures. The rate of cleavage of d(G-A-A-C-C-G-G-A-G-A) is several-fold faster than that of d(T-C-T-C-C-G-G-T-T). Single strand phi X174 DNA is also, cleaved by Msp I endonuclease giving well defined fragments. 5'-Nucleotide analysis of the fragments generated from single strand and replicating form DNA suggest that cleavage occurs at the recognition sequence d(C-C-G-G). The data show that Msp I endonuclease cleaves single strand oligonucleotides and prefers a recognition sequence surrounded by purine nucleotides. A general model for endonuclease cleavage of single strand and duplex DNA is presented.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Bacteriophage phi X 174 / analysis*
  • DNA Restriction Enzymes / metabolism*
  • DNA, Single-Stranded*
  • DNA, Viral*
  • Deoxyribonuclease HpaII
  • Kinetics
  • Oligodeoxyribonucleotides / analysis*
  • Oligonucleotides / analysis*

Substances

  • DNA, Single-Stranded
  • DNA, Viral
  • Oligodeoxyribonucleotides
  • Oligonucleotides
  • DNA Restriction Enzymes
  • Deoxyribonuclease HpaII