Obtaining the recombinant fusion protein OprF-aTox of <i>Pseudomonas aeruginosa</i> whith protective properties
Into the cells of Escherichia coli (strain BL21(DE3)) two forms of the fusion protein Pseudomonas aeruginosa containing the full length outer membrane protein F (OprF) and nontoxic form of Exotoxin A (without 106 C-terminal amino acid residues) have been synthesized. Two recombinant genes were inser...
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| Format: | Article |
| Language: | Russian |
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Ministry of Health of the Russian Federation. Federal State Budgetary Institution «Scientific Centre for Expert Evaluation of Medicinal Products»
2018-02-01
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| Series: | Биопрепараты: Профилактика, диагностика, лечение |
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| Online Access: | https://www.biopreparations.ru/jour/article/view/48 |
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| author | A. V. Soldatenkova N. A. Mihailova E. M. Zimina E. I. Leonova A. A. Kaloshin |
| author_facet | A. V. Soldatenkova N. A. Mihailova E. M. Zimina E. I. Leonova A. A. Kaloshin |
| author_sort | A. V. Soldatenkova |
| collection | DOAJ |
| description | Into the cells of Escherichia coli (strain BL21(DE3)) two forms of the fusion protein Pseudomonas aeruginosa containing the full length outer membrane protein F (OprF) and nontoxic form of Exotoxin A (without 106 C-terminal amino acid residues) have been synthesized. Two recombinant genes were inserted into plasmid pET28 in different order: oprF-atox and atox-oprF. Only oprF-atox variant allowed to obtain the recombinant protein sufficient for purification by affinity chromatography on Ni-Sepharose. The recombinant fusion protein OprF-aTox showed a high specificity in interaction with the preparations of polyclonal immune rabbit serum to the recombinant OprF, the recombinant nontoxic form of Exotoxin A and the bacterial cells of P. aeruginosa. The purified recombinant fusion protein OprF-aTox after two immunizations protected the mice against toxigenic strain of P. aeruginosa (РА-103) being injected intraperitoneally. The index of efficiency of protective properties of OprF-aTox in the optimal dose (50 µg protein per mouse) was 3.5. This was more efficient than in cases when the recombinant OprF and the recombinant toxoid were injected separately. The indexes of efficiency of protective properties of OprF and the toxoid were 2.1 and 2.0. |
| format | Article |
| id | doaj-art-b285aaf19e234dc2b8d9a0b617dfc9d0 |
| institution | Kabale University |
| issn | 2221-996X 2619-1156 |
| language | Russian |
| publishDate | 2018-02-01 |
| publisher | Ministry of Health of the Russian Federation. Federal State Budgetary Institution «Scientific Centre for Expert Evaluation of Medicinal Products» |
| record_format | Article |
| series | Биопрепараты: Профилактика, диагностика, лечение |
| spelling | doaj-art-b285aaf19e234dc2b8d9a0b617dfc9d02025-08-20T03:43:15ZrusMinistry of Health of the Russian Federation. Federal State Budgetary Institution «Scientific Centre for Expert Evaluation of Medicinal Products»Биопрепараты: Профилактика, диагностика, лечение2221-996X2619-11562018-02-011629610048Obtaining the recombinant fusion protein OprF-aTox of <i>Pseudomonas aeruginosa</i> whith protective propertiesA. V. Soldatenkova0N. A. Mihailova1E. M. Zimina2E. I. Leonova3A. A. Kaloshin4I. Mechnikov Research Institute of Vaccines and SeraI. Mechnikov Research Institute of Vaccines and SeraI. Mechnikov Research Institute of Vaccines and SeraI. Mechnikov Research Institute of Vaccines and SeraI. Mechnikov Research Institute of Vaccines and SeraInto the cells of Escherichia coli (strain BL21(DE3)) two forms of the fusion protein Pseudomonas aeruginosa containing the full length outer membrane protein F (OprF) and nontoxic form of Exotoxin A (without 106 C-terminal amino acid residues) have been synthesized. Two recombinant genes were inserted into plasmid pET28 in different order: oprF-atox and atox-oprF. Only oprF-atox variant allowed to obtain the recombinant protein sufficient for purification by affinity chromatography on Ni-Sepharose. The recombinant fusion protein OprF-aTox showed a high specificity in interaction with the preparations of polyclonal immune rabbit serum to the recombinant OprF, the recombinant nontoxic form of Exotoxin A and the bacterial cells of P. aeruginosa. The purified recombinant fusion protein OprF-aTox after two immunizations protected the mice against toxigenic strain of P. aeruginosa (РА-103) being injected intraperitoneally. The index of efficiency of protective properties of OprF-aTox in the optimal dose (50 µg protein per mouse) was 3.5. This was more efficient than in cases when the recombinant OprF and the recombinant toxoid were injected separately. The indexes of efficiency of protective properties of OprF and the toxoid were 2.1 and 2.0.https://www.biopreparations.ru/jour/article/view/48pseudomonas aeruginosaбелок f наружной мембраныэкзотоксин аанатоксинслитый рекомбинантный белок oprf-атохиммунизацияpseudomonas aeruginosaouter membrane protein fexotoxin atoxoidrecombinant fusion protein oprf-atoximmunization |
| spellingShingle | A. V. Soldatenkova N. A. Mihailova E. M. Zimina E. I. Leonova A. A. Kaloshin Obtaining the recombinant fusion protein OprF-aTox of <i>Pseudomonas aeruginosa</i> whith protective properties Биопрепараты: Профилактика, диагностика, лечение pseudomonas aeruginosa белок f наружной мембраны экзотоксин а анатоксин слитый рекомбинантный белок oprf-атох иммунизация pseudomonas aeruginosa outer membrane protein f exotoxin a toxoid recombinant fusion protein oprf-atox immunization |
| title | Obtaining the recombinant fusion protein OprF-aTox of <i>Pseudomonas aeruginosa</i> whith protective properties |
| title_full | Obtaining the recombinant fusion protein OprF-aTox of <i>Pseudomonas aeruginosa</i> whith protective properties |
| title_fullStr | Obtaining the recombinant fusion protein OprF-aTox of <i>Pseudomonas aeruginosa</i> whith protective properties |
| title_full_unstemmed | Obtaining the recombinant fusion protein OprF-aTox of <i>Pseudomonas aeruginosa</i> whith protective properties |
| title_short | Obtaining the recombinant fusion protein OprF-aTox of <i>Pseudomonas aeruginosa</i> whith protective properties |
| title_sort | obtaining the recombinant fusion protein oprf atox of i pseudomonas aeruginosa i whith protective properties |
| topic | pseudomonas aeruginosa белок f наружной мембраны экзотоксин а анатоксин слитый рекомбинантный белок oprf-атох иммунизация pseudomonas aeruginosa outer membrane protein f exotoxin a toxoid recombinant fusion protein oprf-atox immunization |
| url | https://www.biopreparations.ru/jour/article/view/48 |
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