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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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