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A secretory system for bacterial production of high-profile protein targets.

ABSTRACT: Escherichia coli represents a robust, inexpensive expression host for the production of recombinant proteins. However, one major limitation is that certain protein classes do not express well in a biologically relevant form using standard expression approaches in the cytoplasm of E. coli. To improve the usefulness of the E. coli expression platform we have investigated combinations of promoters and selected N-terminal fusion tags for the extracellular expression of human target proteins. A comparative study was conducted on 24 target proteins fused to outer membrane protein A (OmpA), outer membrane protein F (OmpF) and osmotically inducible protein Y (OsmY). Based on the results of this initial study, we carried out an extended expression screen employing the OsmY fusion and multiple constructs of a more diverse set of human proteins. Using this high-throughput compatible system, we clearly demonstrate that secreted biomedically relevant human proteins can be efficiently retrieved and purified from the growth medium.

SUBMITTER: Kotzsch A

PROVIDER: S-EPMC3064838 | biostudies-literature | 2011 Mar

REPOSITORIES: biostudies-literature

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A secretory system for bacterial production of high-profile protein targets.

Kotzsch Alexander A Vernet Erik E Hammarström Martin M Berthelsen Jens J Weigelt Johan J Gräslund Susanne S Sundström Michael M

Protein science : a publication of the Protein Society 20110301 3

Escherichia coli represents a robust, inexpensive expression host for the production of recombinant proteins. However, one major limitation is that certain protein classes do not express well in a biologically relevant form using standard expression approaches in the cytoplasm of E. coli. To improve the usefulness of the E. coli expression platform we have investigated combinations of promoters and selected N-terminal fusion tags for the extracellular expression of human target proteins. A compa ...[more]

PMID: 21308845

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A secretory system for bacterial production of high-profile protein targets.

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A secretory system for bacterial production of high-profile protein targets.

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