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Biochemistry (Moscow)

, Volume 83, Issue 3, pp 259–269 | Cite as

Expression of Soluble Active Interferon αA in Escherichia coli Periplasm by Fusion with Thermostable Lichenase Using the Domain Insertion Approach

  • A. A. Tyurin
  • K. V. Kabardaeva
  • O. N. Mustafaev
  • O. S. Pavlenko
  • N. S. Sadovskaya
  • V. S. Fadeev
  • E. A. Zvonova
  • I. V. Goldenkova-PavlovaEmail author
Article

Abstract

A recombinant DNA in which the interferon αA (IFN–αA) gene sequence is integrated into a loop region of the gene coding thermostable lichenase was constructed. This approach of insertion fusion with thermostable lichenase is advantageous in terms of increasing the solubility, stability, and production of the fusion partner in soluble form in general and in the periplasm of bacterial cells in particular. Thus, the insertion of IFN–αA into the loop (53 a.a.) of thermostable lichenase from Clostridium thermocellum resulted in effective expression of the soluble form of the recombinant protein in the periplasm of Escherichia coli without any compromise in biological activity of IFN–αA, while the thermostable lichenase retained its ability for functional folding without dramatic loss of its basic activity and thermostability.

Keywords

interferon αA thermostable lichenase domain insertion expression solubility biological activity 

Abbreviations

CPE

cytopathogenic effect

FBS

fetal bovine serum

IFN

interferon

LicB

thermostable lichenase

MBP

maltose–binding protein

P–EP

protein preparation after ethanol precipitation

PP

purified protein

TPL

total protein lysate

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Copyright information

© Pleiades Publishing, Ltd. 2018

Authors and Affiliations

  • A. A. Tyurin
    • 1
    • 2
  • K. V. Kabardaeva
    • 1
    • 2
  • O. N. Mustafaev
    • 3
  • O. S. Pavlenko
    • 1
    • 2
  • N. S. Sadovskaya
    • 1
  • V. S. Fadeev
    • 1
  • E. A. Zvonova
    • 2
  • I. V. Goldenkova-Pavlova
    • 1
    Email author
  1. 1.Institute of Plant PhysiologyRussian Academy of SciencesMoscowRussia
  2. 2.Department of Genetics and BiotechnologyRussian State Agrarian University − Moscow Timiryazev Agricultural AcademyMoscowRussia
  3. 3.Department of Biophysics and Molecular BiologyBaku State UniversityBakuAzerbaijan

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