Rational engineering of type II restriction endonuclease DNA binding and cleavage specificity.

ABSTRACT: The type II restriction endonucleases are indispensible tools for molecular biology. Although enzymes recognizing nearly 300 unique sequences are known, the ability to engineer enzymes to recognize any sequence of choice would be valuable. However, previous attempts to engineer new recognition specificity have met limited success. Here we report the rational engineering of multiple new type II specificities. We recently identified a family of MmeI-like type II endonucleases that have highly similar protein sequences but different recognition specificity. We identified the amino-acid positions within these enzymes that determine position specific DNA base recognition at three positions within their recognition sequences through correlations between their aligned amino-acid residues and alig

SUBMITTER: Morgan RD 

PROVIDER: S-EPMC2731914 | biostudies-literature | 2009 Aug

REPOSITORIES: biostudies-literature