Cloning, expression, purification, crystallization and initial crystallographic analysis of transcription factor DksA from Escherichia coli.
The Escherichia coli gene encoding a regulator of stringent response and virulence, DksA, which contains a canonical Zn finger motif, was cloned and expressed, and the purified protein was crystallized by the hanging-drop vapor-diffusion technique in two different space groups, P2(1)2(1)2(1) (a = 91.32, b = 96.59, c = 117.48 A) and C222 (a = 80.6, b = 115.1, c = 149.57 A). The crystals belonging to space group P2(1)2(1)2(1), improved by macroseeding, diffract beyond 2.0 A at a synchrotron. Three complete atomic resolution multiple anomalous dispersion diffraction data sets were collected from the same crystal of the P2(1)2(1)2(1) crystal form at the absorption edge for Zn atoms.
Vassylyeva
Marina N
MN
Genomic Sciences Center, RIKEN Yokohama Institute, 1-7-22 Suehiro-cho, Tsurumi Yokohama 230-0045, Japan.
Perederina
Anna A
AA
Svetlov
Vladimir
V
Yokoyama
Shigeyuki
S
Artsimovitch
Irina
I
Vassylyev
Dmitry G
DG
D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000737:chemistry* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D008958:Models, Molecular; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017356:Synchrotrons; D016335:Zinc Fingers
D029968:Escherichia coli Proteins; C090751:dksA protein, E coli