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Recombinant fluorescent models for studying the diphtheria toxin

Abstract

Diphtheria toxin is a major factor of the pathogenicity of the causative agent of diphtheria Corynebacterium. Due to a small size, it is of considerable interest as the basis for the development of synthetic protein molecules with a transport function, e.g., immunotoxins. In this work we describe the expression and characterized nontoxic recombinant fluorescent derivatives of the diphtheria toxin and its nontoxic CRM197 mutant. The proteins obtained can be used for studying receptor-binding and transport functions of the toxin in cells, evaluation of the expression level of the toxin proHB-EGF receptor membranes, immunization and generation of specific antibodies against the toxin, as well as for the development of diagnostic test-systems for the diphtheria toxin and antitoxic antibodies.

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Abbreviations

DT:

diphtheria toxin

BSA:

bovine serum albumin

DABCO:

1,4-diazobicyclooctane

IPTG:

isopropyl-β-D-1-thiogalactopyranoside

LD:

a lethal dose

PBS:

phosphate buffered saline

proHB-EGF:

precursor of the heparin-binding epidermal growth factor-like growth factor

PVA:

polyvinyl alcohol

Rd:

a diphtheria toxin R domain

SbA and SbB:

diphtheria toxin A and B subunits

SOE-PCR:

splicing with overlap extension PCR

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Correspondence to A. J. Labyntsev.

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Original Russian Text © A.J. Labyntsev, N.V. Korotkevych, K.J. Manoilov, A.A. Kaberniuk, D.V. Kolybo, S.V. Komisarenko, 2014, published in Bioorganicheskaya Khimiya, 2014, Vol. 40, No. 4, pp. 433–442.

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Labyntsev, A.J., Korotkevych, N.V., Manoilov, K.J. et al. Recombinant fluorescent models for studying the diphtheria toxin. Russ J Bioorg Chem 40, 401–409 (2014). https://doi.org/10.1134/S1068162014040086

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  • DOI: https://doi.org/10.1134/S1068162014040086

Keywords

  • fluorescent proteins
  • molecular cloning
  • proHB-EGF
  • recombinant nontoxic bacterial toxin analogues