Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of the regulator AcrR from Escherichia coli

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Nov 1;62(Pt 11):1150-2. doi: 10.1107/S1744309106042576. Epub 2006 Oct 20.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Base Sequence
  • Cloning, Molecular
  • Crystallization
  • DNA Primers
  • Escherichia coli / genetics*
  • Escherichia coli Proteins / chemistry*
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / isolation & purification
  • Molecular Sequence Data
  • Repressor Proteins / chemistry*
  • Repressor Proteins / genetics
  • Repressor Proteins / isolation & purification
  • X-Ray Diffraction

Substances

  • AcrR protein, E coli
  • DNA Primers
  • Escherichia coli Proteins
  • Repressor Proteins