HEAT-RESISTANT FLAP ENDONUCLEASE ORIGINATING FROM HYPERTHERMOPHILIC BACTERIUM BELONGING TO GENUS PYROCOCCUS

摘要

PROBLEM TO BE SOLVED: To provide the subject new enzyme originating from hyperthermophilic bacterium, consisting of a thermophilic Flap endonuclease having a specific amino acid sequence and high optimum temperature and effective for gene recombination, gene shuffling, etc., based on low homology. ;SOLUTION: This protein is a new thermophilic Flap endonuclease composed of the amino acid sequence of the formula and having an optimum temperature of ≥75°C or a new protein composed of the amino acid sequence of the formula provided that one or several amino acids are depleted, substituted or added and having Flap endonuclease activity. The protein is useful for the gene recombination, gene shuffling, etc., based on low homology. The thermophilic Flap endonuclease can be produced by carrying out PCR reaction with a primer using the chromosome DNA of Pyrococcus horikoshi JCM9974 as a template, integrating the obtained gene into a protein expression plasmid and culturing its transformed E.coli in a medium.;COPYRIGHT: (C)1999,JPO