Expression, purification and crystallization of the cell-division protein YgfE from Escherichia coli

Stephen G. Addinall, Kenneth A. Johnson, Timothy Dafforn, Corinne Smith, Alison Rodger, Raul Paco Gomez, Katherine Sloan, Anne Blewett, David J. Scott, David I. Roper*

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

An open reading frame designated b2910 (ygfE) in the Escherichia coli K12-MG1655 genome sequence, identified as a possible homologue to the cell-division protein ZapA, was cloned into the high-expression plasmid pETDuet-1 and overexpressed in E. coli BL21 (DES)-AI. The protein was purified in three steps to 99% purity. Crystals were obtained by the hanging-drop vapour-diffusion method at 291 K from a wide range of screened conditions, but principally from solutions containing 0.1 M HEPES pH 7.0, 18% PEG 6000, 5 mM CaCl2. Diffraction data to 1.8 Å were collected at the European Synchrotron Radiation Facility (ESRF). The crystals belong to space group P6122 or P6522, with unit-cell parameters a = 53.8, b = 53.8, c = 329.7 Å, α = β = 90, γ = 120°.

Original languageEnglish
Pages (from-to)305-307
Number of pages3
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
Volume61
Issue number3
DOIs
Publication statusPublished - 2005
Externally publishedYes

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