Open AccessOverexpression, purification, crystallization and preliminary X‐ray studies of Vibrio cholerae EpsG
Author(s) -
Jens Jason,
Raghunathan Kannan,
Vago Frank,
Arvidson Dennis
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309109017163
Subject(s) - vibrio cholerae , escherichia coli , recombinant dna , crystallization , plasmid , enterotoxin , microbiology and biotechnology , chemistry , resolution (logic) , secretion , biology , zinc , vibrio , bacteria , biochemistry , gene , genetics , organic chemistry , artificial intelligence , computer science
EpsG is the major pseudopilin protein of the Vibrio cholerae type II secretion system. An expression plasmid that encodes an N‐terminally truncated form of EpsG with a C‐terminal noncleavable His tag was constructed. Recombinant EpsG was expressed in Escherichia coli ; the truncated protein was purified and crystallized by hanging‐drop vapor diffusion against a reservoir containing 6 m M zinc sulfate, 60 m M MES pH 6.5, 15% PEG MME 550. The crystals diffracted X‐rays to a resolution of 2.26 Å and belonged to space group P 2 1 , with unit‐cell parameters a = 88.61, b = 70.02, c = 131.54 Å.