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

A. A. Tyurin1,2, K. V. Kabardaeva1,2, O. N. Mustafaev3, O. S. Pavlenko1,2, N. S. Sadovskaya1, V. S. Fadeev1, E. A. Zvonova2, and I. V. Goldenkova-Pavlova1*

1Institute of Plant Physiology, Russian Academy of Sciences, 127276 Moscow, Russia; E-mail: irengold58@gmail.com

2Russian State Agrarian University – Moscow Timiryazev Agricultural Academy, Department of Genetics and Biotechnology, 127550 Moscow, Russia; E-mail: alexjofar@gmail.com

3Baku State University, Department of Biophysics and Molecular Biology, Baku, AZ 1148, Azerbaijan; E-mail: orkhan@bioset.org

* To whom correspondence should be addressed.

Received August 30, 2017; Revision received November 2, 2017
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.
KEY WORDS: interferon αA, thermostable lichenase, domain insertion, expression, solubility, biological activity

DOI: 10.1134/S0006297918030069