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Improved production of membrane proteins in Escherichia coli by selective codon substitutions
Author(s) -
Nørholm Morten H.H.,
Toddo Stephen,
Virkki Minttu T.I.,
Light Sara,
von Heijne Gunnar,
Daley Daniel O.
Publication year - 2013
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/j.febslet.2013.05.063
Subject(s) - escherichia coli , chemistry , escherichia coli proteins , membrane protein , biochemistry , membrane , biology , gene
Membrane proteins are extremely challenging to produce in sufficient quantities for biochemical and structural analysis and there is a growing demand for solutions to this problem. In this study we attempted to improve expression of two difficult‐to‐express coding sequences ( araH and narK ) for membrane transporters. For both coding sequences, synonymous codon substitutions in the region adjacent to the AUG start led to significant improvements in expression, whereas multi‐parameter sequence optimization of codons throughout the coding sequence failed. We conclude that coding sequences can be re‐wired for high‐level protein expression by selective engineering of the 5′ coding sequence with synonymous codons, thus circumventing the need to consider whole sequence optimization.