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Collection and characterisation of bacterial membrane proteins

Saidijam, Massoud, Psakis, Georgios, Clough, J.L., Meuller, J, Suzuki, S, Hoyle, C.J., Palmer, S.L., Morrison, S.M., Pos, M.K., Essenberg, R.C., Maiden, M.C.J., Abu-bakr, A, Baumberg, S.G., Neyfakh, A.A., Griffith, J.K., Stark, M.J., Ward, A, O'Reilly, J, Rutherford, N.G., Phillips-Jones, M.K. and Henderson, P.J.F. (2003) Collection and characterisation of bacterial membrane proteins. FEBS Letters, 555 (1). pp. 170-175. ISSN 0014-5793

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A general strategy for the amplified expression in Escherichia coli of membrane transport and receptor proteins from other bacteria is described. As an illustration we report the cloning of the putative alpha-ketoglutarate membrane transport gene from the genome of Helicobacter pylori, overexpression of the protein tagged with RGS(His)6 at the C-terminus, and its purification in mg quantities. The retention of structural and functional integrity was verified by circular dichroism spectroscopy and reconstitution of transport activity. This strategy for overexpression and purification is extended to additional membrane proteins from H. pylori and from other bacteria.

Item Type: Article
Subjects: Q Science > Q Science (General)
Q Science > QC Physics
Q Science > QD Chemistry
Schools: School of Applied Sciences
Depositing User: Georgios Psakis
Date Deposited: 05 Sep 2013 11:14
Last Modified: 05 Sep 2013 11:14


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