Open AccessCrystallization and preliminary X‐ray analysis of flap endonuclease 1 (FEN1) from Desulfurococcus amylolyticus
Author(s) -
Mase Tomoko,
Kubota Keiko,
Miyazono Kenichi,
Kawarabayasi Yutaka,
Tanokura Masaru
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309109031248
Subject(s) - okazaki fragments , crystallization , nuclease , crystallography , dna , endonuclease , chemistry , dna replication , biology , microbiology and biotechnology , biochemistry , eukaryotic dna replication , organic chemistry
Flap endonuclease 1 (FEN1) is a structure‐specific nuclease that removes 5′‐overhanging flaps in DNA repair and removes the RNA/DNA primer during maturation of the Okazaki fragment in lagging‐strand DNA replication. FEN1 from the hyperthermophilic archaeon Desulfurococcus amylolyticus was expressed in Escherichia coli , purified and crystallized using the sitting‐drop vapour‐diffusion method with monoammonium dihydrogen phosphate as the precipitant at pH 8.3. X‐ray diffraction data were collected to 2.00 Å resolution. The space group of the crystal was determined as the primitive hexagonal space group P 321, with unit‐cell parameters a = b = 103.76, c = 84.58 Å. The crystal contained one molecule in the asymmetric unit.