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1. (WO2003060152) ALTERATION OF RESTRICTION ENDONUCLEASE SPECIFICITY BY GENETIC SELECTION

Pub. No.:    WO/2003/060152    International Application No.:    PCT/US2003/000542
Publication Date: Jul 24, 2003 International Filing Date: Jan 9, 2003
IPC: C12N 9/22
C12N 15/10
Applicants: NEW ENGLAND BIOLABS, INC.
SAMUELSON, James, C.
XU, Shuang-yong
Inventors: SAMUELSON, James, C.
XU, Shuang-yong
Title: ALTERATION OF RESTRICTION ENDONUCLEASE SPECIFICITY BY GENETIC SELECTION
Abstract:
Methods and compositions are provided for altering the DNA recognition and cleavage characteristics of an endonuclease without prior knowledge of the endonuclease's three-dimensional structure and/or amino acid residues responsible for activity and/or specificity. Methods include subjecting a mutagenized endonuclease gene library to a genetic selection in prokaryotic cells which tolerate the expression of mutated endonuclease and where the endonuclease is active and determining the altered recognition-site specificity for the endonuclease.