Premium
Collection and characterisation of bacterial membrane proteins
Author(s) -
Saidijam Massoud,
Psakis Georgios,
Clough Joanne L,
Meuller Johan,
Suzuki Shun'ichi,
Hoyle Christopher J,
Palmer Sarah L,
Morrison Scott M,
Pos Martin K,
Essenberg Richard C,
Maiden Martin C.J,
Abu-bakr Atif,
Baumberg Simon G,
Neyfakh Alex A,
Griffith Jeffrey K,
Stark Michael J,
Ward Alison,
O'Reilly John,
Rutherford Nicholas G,
Phillips-Jones Mary K,
Henderson Peter J.F
Publication year - 2003
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(03)01148-7
Subject(s) - escherichia coli , bacteria , cloning (programming) , membrane protein , bacterial outer membrane , membrane transport , biochemistry , transport protein , chemistry , membrane , biology , gene , microbiology and biotechnology , genetics , computer science , programming language
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 α‐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.