Localization and activity of recombinant streptavidin in cell fractions of Escherichia coli



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Abstract

The authors provide a genetic construct for obtaining recombinant biologically active wild type streptavidin with high yield. Addition of leader peptide and oligohistidine sequence to the streptavidin sequence makes it possible to isolate soluble streptavidin from the culture medium and from the soluble fraction. Temperature conditions for inducing of protein synthesis were found to have a significant effect on the distribution profile of streptavidin between fractions. The obtained protein product is not contaminated with endogenous biotin. The provided method excludes denaturation and renaturation steps which are necessary for isolation of the desired product from inclusion bodies but substantially reduce the yield. Thus, the provided approach allows to increase the yield of biologically highly active strepatividin which is capable of binding biotin and biotin-containing compounds and can be used for various practical purposes.

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About the authors

D V Zotova

Institute of Experimental Medicine

N A Grudinina

Institute of Experimental Medicine

O I Antimonova

Institute of Experimental Medicine

M M Shavlovsky

North-Western State Medical University named after I.I. Mechnikov

D S Polyakov

Institute of Experimental Medicine

References

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Copyright (c) 2018 Zotova D.V., Grudinina N.A., Antimonova O.I., Shavlovsky M.M., Polyakov D.S.

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