Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of the regulator AcrR from Escherichia coli.
This paper describes the cloning, expression, purification and preliminary X-ray data analysis of the AcrR regulatory protein. The Escherichia coli AcrR is a member of the TetR family of transcriptional regulators. It regulates the expression of the AcrAB multidrug transporter. Recombinant AcrR with a 6xHis tag at the C-terminus was expressed in E. coli and purified by metal-affinity chromatography. The protein was crystallized using hanging-drop vapor diffusion. X-ray diffraction data were collected from cryocooled crystals at a synchrotron light source. The best crystal diffracted to 2.5 A. The space group was determined to be P3(2), with unit-cell parameters a = b = 46.61, c = 166.16 A.
Li
Ming
M
Department of Physics and Astronomy, Iowa State University, Ames, IA 50011, USA.
Qiu
Xi
X
Su
Chih-Chia
CC
Long
Feng
F
Gu
Ruoyu
R
McDermott
Gerry
G
Yu
Edward W
EW
D001483:Base Sequence; D003001:Cloning, Molecular; D003460:Crystallization; D017931:DNA Primers; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D012097:Repressor Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D014961:X-Ray Diffraction
C515217:AcrR protein, E coli; D017931:DNA Primers; D029968:Escherichia coli Proteins; D012097:Repressor Proteins