Collection and characterisation of bacterial membrane proteins
Open Access
- 17 October 2003
- journal article
- Published by Wiley in FEBS Letters
- Vol. 555 (1) , 170-175
- https://doi.org/10.1016/s0014-5793(03)01148-7
Abstract
A general strategy for the amplified expression inEscherichia coliof membrane transport and receptor proteins from other bacteria is described. As an illustration we report the cloning of the putative α‐ketoglutarate membrane transport gene from the genome ofHelicobacter pylori, overexpression of the protein tagged with RGS(His)6at 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 fromH. pyloriand from other bacteria.Keywords
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