Open AccessCloning, expression, purification, crystallization and preliminary X‐ray diffraction analysis of the regulator AcrR from Escherichia coli
Author(s) -
Li Ming,
Qiu Xi,
Su ChihChia,
Long Feng,
Gu Ruoyu,
McDermott Gerry,
Yu Edward W.
Publication year - 2006
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309106042576
Subject(s) - escherichia coli , cloning (programming) , recombinant dna , microbiology and biotechnology , tetr , chemistry , affinity chromatography , biology , crystallization , gene expression , gene , biochemistry , repressor , enzyme , organic chemistry , computer science , programming language
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 6×His 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 Å. The space group was determined to be P 3 2 , with unit‐cell parameters a = b = 46.61, c = 166.16 Å.