Director-general of Agency of Industrial Science and Technology

National Institute of Advanced Industrial Science and Technology

Eriko Matsui

Ikuo Matsui

Kazuhiko Ishikawa

Yoshitsugu Kosugi

Satoko Kawasaki

A mutation is introduced into the substrate-binding site of flap endonuclease to prepare a mutant with modified substrate specificity. Using the mutant as a reagent for the analysis of genetic polymorphism, the analysis of genetic polymorphism can be performed more accurately, easily and sensitively as compared with conventional methods.

Flap endonuclease mutants

The present invention relates to a thermostable Flap endonuclease whose optimum temperature is 75&deg; C. or more and DNA coding for (a) a protein consisting of the amino acid sequence shown in SEQ ID NO:2 or (b) a protein with Flap endonuclease activity, consisting of an amino acid sequence where in the amino acid sequence (a), one or more amino acids are deleted, substituted or added. According to the present invention, there is provided a thermostable Flap endonuclease whose optimum temperature for reaction is 75&deg; C. or more. Further, this enzyme is thermally stable, so it becomes possible to develop new techniques of conducting artificial homologous recombination or genetic shuffling highly efficiently by coupling the enzyme reaction with PCR.

Thermostable flap endonuclease derived from hyperthermophile bacterium belonging to the genus pyrococcus

Thermostable flap endonuclease derived from a hyperthermophile bacterium belonging to the genus pyrococcus

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Ibaraki, Japan

Eriko Matsui