Open AccessPurification, crystallization, X‐ray diffraction analysis and phasing of an engineered single‐chain Pvu II restriction endonuclease
Author(s) -
Meramveliotaki Chrysi,
Kotsifaki Dina,
Androulaki Maria,
Hountas Athanasios,
Eliopoulos Elias,
Kokkinidis Michael
Publication year - 2007
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309107040377
Subject(s) - restriction enzyme , crystallization , endonuclease , x ray crystallography , diffraction , crystallography , phaser , materials science , microbiology and biotechnology , chemistry , dna , biology , genetics , physics , optics , organic chemistry
The restriction endonuclease Pvu II from Proteus vulgaris has been converted from its wild‐type homodimeric form into the enzymatically active single‐chain variant sc Pvu II by tandemly joining the two subunits through the peptide linker Gly‐Ser‐Gly‐Gly. sc Pvu II, which is suitable for the development of programmed restriction endonucleases for highly specific DNA cleavage, was purified and crystallized. The crystals diffract to a resolution of 2.35 Å and belong to space group P 4 2 , with unit‐cell parameters a = b = 101.92, c = 100.28 Å and two molecules per asymmetric unit. Phasing was successfully performed by molecular replacement.