Class-IIS restriction enzymes--a review

Gene. 1991 Apr:100:13-26. doi: 10.1016/0378-1119(91)90345-c.


Class-IIS restriction enzymes (ENases-IIS) interact with two discrete sites on double-stranded DNA: the recognition site, which is 4-7 bp long, and the cleavage site, usually 1-20 bp away from the recognition site. The recognition sequences of ENases-IIS are totally (or partially) asymmetric and all of the characterized ENases-IIS are monomeric. A total of 35 ENases-IIS are described (80, if all isoschizomers are taken into consideration) together with ten related ENases (class IIT), and 15 cognate methyltransferases (MTases-IIS). The physical, chemical, and molecular properties of the ENases-IIS and MTases-IIS are reviewed and many unique applications of this class of enzymes are described, including: precise trimming of DNA; retrieval of cloned fragments; gene assembly; use as a universal restriction enzyme; cleavage of single-stranded DNA; detection of point mutations; tandem amplification; printing-amplification reaction; and localization of methylated bases.

Publication types

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

MeSH terms

  • Base Sequence
  • DNA / genetics
  • Deoxyribonucleases, Type II Site-Specific / metabolism*
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • Restriction Mapping
  • Substrate Specificity


  • DNA
  • Deoxyribonucleases, Type II Site-Specific