Expression of Soluble Active Interferon αA in Escherichia coli Periplasm by Fusion with Thermostable Lichenase Using the Domain Insertion Approach
- Authors: Tyurin A.A.1,2, Kabardaeva K.V.1,2, Mustafaev O.N.3, Pavlenko O.S.1,2, Sadovskaya N.S.1, Fadeev V.S.1, Zvonova E.A.2, Goldenkova-Pavlova I.V.1
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Affiliations:
- Institute of Plant Physiology
- Department of Genetics and Biotechnology
- Department of Biophysics and Molecular Biology
- Issue: Vol 83, No 3 (2018)
- Pages: 259-269
- Section: Article
- URL: https://journals.rcsi.science/0006-2979/article/view/151614
- DOI: https://doi.org/10.1134/S0006297918030069
- ID: 151614
Cite item
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.
About the authors
A. A. Tyurin
Institute of Plant Physiology; Department of Genetics and Biotechnology
Email: irengold58@gmail.com
Russian Federation, Moscow, 127276; Moscow, 127550
K. V. Kabardaeva
Institute of Plant Physiology; Department of Genetics and Biotechnology
Email: irengold58@gmail.com
Russian Federation, Moscow, 127276; Moscow, 127550
O. N. Mustafaev
Department of Biophysics and Molecular Biology
Email: irengold58@gmail.com
Azerbaijan, Baku, AZ, 1148
O. S. Pavlenko
Institute of Plant Physiology; Department of Genetics and Biotechnology
Email: irengold58@gmail.com
Russian Federation, Moscow, 127276; Moscow, 127550
N. S. Sadovskaya
Institute of Plant Physiology
Email: irengold58@gmail.com
Russian Federation, Moscow, 127276
V. S. Fadeev
Institute of Plant Physiology
Email: irengold58@gmail.com
Russian Federation, Moscow, 127276
E. A. Zvonova
Department of Genetics and Biotechnology
Email: irengold58@gmail.com
Russian Federation, Moscow, 127550
I. V. Goldenkova-Pavlova
Institute of Plant Physiology
Author for correspondence.
Email: irengold58@gmail.com
Russian Federation, Moscow, 127276