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Expression of G-protein coupled receptors in Escherichia coli for structural studies

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Abstract

To elaborate a high-performance system for expression of genes of G-protein coupled receptors (GPCR), methods of direct and hybrid expression of 17 GPCR genes in Escherichia coli and selection of strains and bacteria cultivation conditions were investigated. It was established that expression of most of the target GPCR fused with the N-terminal fragment of OmpF or Mistic using media for autoinduction provides high output (up to 50 mg/liter).

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Abbreviations

GPCR:

G-protein coupled receptors

GST:

glutathione-S-transferase

IPTG:

isopropyl-β-D-thiogalactoside

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Authors and Affiliations

  1. Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, 117997, Moscow, Russia

    L. E. Petrovskaya, A. A. Shulga, O. V. Bocharova, Ya. S. Ermolyuk, E. A. Kryukova, V. V. Chupin, A. S. Arseniev & M. P. Kirpichnikov

  2. Novartis Institutes for BioMedical Research, Basel, Switzerland

    M. J. J. Blommers

  3. Faculty of Biology, Lomonosov Moscow State University, 119991, Moscow, Russia

    M. P. Kirpichnikov

Authors
  1. L. E. Petrovskaya
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  2. A. A. Shulga
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  3. O. V. Bocharova
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  4. Ya. S. Ermolyuk
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  5. E. A. Kryukova
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  6. V. V. Chupin
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  7. M. J. J. Blommers
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  8. A. S. Arseniev
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  9. M. P. Kirpichnikov
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Corresponding author

Correspondence to L. E. Petrovskaya.

Additional information

Original Russian Text © L. E. Petrovskaya, A. A. Shulga, O. V. Bocharova, Ya. S. Ermolyuk, E. A. Kryukova, V. V. Chupin, M. J. J. Blommers, A. S. Arseniev, M. P. Kirpichnikov, 2010, published in Biokhimiya, 2010, Vol. 75, No. 7, pp. 1001–1013.

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Petrovskaya, L.E., Shulga, A.A., Bocharova, O.V. et al. Expression of G-protein coupled receptors in Escherichia coli for structural studies. Biochemistry Moscow 75, 881–891 (2010). https://doi.org/10.1134/S0006297910070102

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  • DOI: https://doi.org/10.1134/S0006297910070102

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