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

doi:10.1107/S1744309105003945

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 journal › Article › peer-review

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 language	English
Pages (from-to)	305-307
Number of pages	3
Journal	Acta Crystallographica Section F: Structural Biology and Crystallization Communications
Volume	61
Issue number	3
DOIs	https://doi.org/10.1107/S1744309105003945
Publication status	Published - 2005
Externally published	Yes

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Addinall, S. G., Johnson, K. A., Dafforn, T., Smith, C., Rodger, A., Gomez, R. P., Sloan, K., Blewett, A., Scott, D. J., & Roper, D. I. (2005). Expression, purification and crystallization of the cell-division protein YgfE from Escherichia coli. Acta Crystallographica Section F: Structural Biology and Crystallization Communications, 61(3), 305-307. https://doi.org/10.1107/S1744309105003945

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

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 {\AA} 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 {\AA}, α = β = 90, γ = 120°.",

author = "Addinall, {Stephen G.} and Johnson, {Kenneth A.} and Timothy Dafforn and Corinne Smith and Alison Rodger and Gomez, {Raul Paco} and Katherine Sloan and Anne Blewett and Scott, {David J.} and Roper, {David I.}",

year = "2005",

doi = "10.1107/S1744309105003945",

language = "English",

volume = "61",

pages = "305--307",

journal = "Acta Crystallographica Section F: Structural Biology and Crystallization Communications",

issn = "1744-3091",

publisher = "Wiley-Blackwell, Wiley",

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Addinall, SG, Johnson, KA, Dafforn, T, Smith, C, Rodger, A, Gomez, RP, Sloan, K, Blewett, A, Scott, DJ & Roper, DI 2005, 'Expression, purification and crystallization of the cell-division protein YgfE from Escherichia coli', Acta Crystallographica Section F: Structural Biology and Crystallization Communications, vol. 61, no. 3, pp. 305-307. https://doi.org/10.1107/S1744309105003945

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AU - Addinall, Stephen G.

AU - Johnson, Kenneth A.

AU - Dafforn, Timothy

AU - Smith, Corinne

AU - Rodger, Alison

AU - Gomez, Raul Paco

AU - Sloan, Katherine

AU - Blewett, Anne

AU - Scott, David J.

AU - Roper, David I.

PY - 2005

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AB - 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°.

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JF - Acta Crystallographica Section F: Structural Biology and Crystallization Communications

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ER -

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

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