Construction and Characterization of a Recombinant Mutant Homolog of the CheW Protein from Thermotoga petrophila RKU-1

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Abstract

In this work, a recombinant chemotaxis CheW protein from Thermotoga petrophila RKU-1 (TpeCheW) and its mutant homologue (TpeCheW-mut) have been obtained. Despite the low homology with the CheW protein of the Escherichia coli bacteria, these proteins do not cause metabolic overload and are well expressed by E. coli laboratory strains. A wide range of features and parameters important for isolation of the TpeCheW-mut protein, such as stability over a wide range of temperatures and pH values, high level of expression, solubility, and the possibility of using simple low-stage purification schemes, including heat pretreatment, has been recognized. Possible directions of using this protein in the practice of scientific and applied research have been formulated and justified.

About the authors

D. V. Grishin

Institute of Biomedical Chemistry

Author for correspondence.
Email: molbiol_ibm@inbox.ru
Russian Federation, ul. Pogodinskaya 10, Moscow, 119121

D. D. Zhdanov

Institute of Biomedical Chemistry

Email: molbiol_ibm@inbox.ru
Russian Federation, ul. Pogodinskaya 10, Moscow, 119121

Ju. A. Gladilina

Institute of Biomedical Chemistry

Email: molbiol_ibm@inbox.ru
Russian Federation, ul. Pogodinskaya 10, Moscow, 119121

V. S. Pokrovsky

Institute of Biomedical Chemistry

Email: molbiol_ibm@inbox.ru
Russian Federation, ul. Pogodinskaya 10, Moscow, 119121

O. V. Podobed

Institute of Biomedical Chemistry

Email: molbiol_ibm@inbox.ru
Russian Federation, ul. Pogodinskaya 10, Moscow, 119121

M. V. Pokrovskaya

Institute of Biomedical Chemistry

Email: molbiol_ibm@inbox.ru
Russian Federation, ul. Pogodinskaya 10, Moscow, 119121

S. S. Aleksandrova

Institute of Biomedical Chemistry

Email: molbiol_ibm@inbox.ru
Russian Federation, ul. Pogodinskaya 10, Moscow, 119121

A. L. Milyushkina

Institute of Biomedical Chemistry

Email: molbiol_ibm@inbox.ru
Russian Federation, ul. Pogodinskaya 10, Moscow, 119121

M. A. Vigovskiy

Institute of Biomedical Chemistry

Email: molbiol_ibm@inbox.ru
Russian Federation, ul. Pogodinskaya 10, Moscow, 119121

N. N. Sokolov

Institute of Biomedical Chemistry

Email: molbiol_ibm@inbox.ru
Russian Federation, ul. Pogodinskaya 10, Moscow, 119121

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