POLUChENIE GIBRIDNOGO BELKA OprF-OprI PSEUDOMONAS AERUGINOSA


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Abstract

Aim. Production of preparation consisting of amino acid sequences of 2 proteins of outer membrane
- OprF and OprI - of P. aeruginosa and study of its protective properties from experimental
P. aeruginosa infection. Materials and methods. Nucleotide sequences coding OprF protein (1
kb) as well as its C-terminal region (0.6 kb) and OprI protein (0.25 kb) were integrated into pQE-
30 plasmid (QIAGEN). And oprF gene (C-terminal region of oprF in variant 2) and oprI gene
were combined and cloned sequentially into a single vector. E. coli M15 strain cells (QIAGEN)
were used for the production of producent strains of recombinant proteins. Protein products were
analyzed by electrophoresis in polyacrylamide gel by Lammle. Purification of recombinant proteins
was performed by affinity chromatography in Ni-sepharose columns. Live virulent culture P.
aeruginosa РА-170015 strain was used for the analysis of protective properties of recombinant
proteins. Results. 2 hybrid recombinant proteins were produced including amino acid sequences
of F and I proteins of outer membrane (OprF and OprI) of P. aeruginosa. Recombinant protein 1
included whole size sequences of OprF and OprI and protein 2 - C-terminal region (including
amino acid residues 192-342) of OprF and whole size sequence of OprI. These recombinant proteins
after 2 immunizations protected mice from the experimental intraperitoneal infection with
P. aeruginosa. Hybrid protein consisting of whole size sequences had the best protective effect.
Conclusion. The results obtained open a perspective for further immunobiological testing of hybrid
recombinant protein OprF-OprI with the aim of creating immunopreparations for prophylaxis of
P. aeruginosa infection.

References

  1. Гатыпова Е.В., Злыгостев С.А., Калошин А.А., Михайлова Н.А. Получение рекомбинантного белка I наружной мембраны (OprI) Pseudomonas aeruginosa и исследование его антигенных свойств. Журн. микробиол. 2008, 6: 50-53.
  2. Руднов В.А. Антибиотикотерапия госпитальных инфекций, вызванных P. aeruginosa. Русс. мед. журн. 2005, 13 (7): 485-490.
  3. Страчунский Л.С., Решедько Г.К., Стецюк О.У. и др. Сравнительная активностьантисинегнойных антибиотиков в отношении нозокомиальных штаммов Pseudomonas aeruginosa, выделенных в отделениях реанимации и интенсивной терапии России. Клинич. микробиол. антимикроб. химиотерап. 2003, 5: 35-46.
  4. Cornelis P. Pseudomonas: Genomics and molecular biology. Caister Academic Press, 2008.
  5. Doring G., Pier G.B. Vaccines and immunotherapy against Pseudomonas aeruginosa. Vaccine. 2008, 26 (8): 1011-1024.
  6. Kaloshin A.A., Gatypova E.V., Mikhailova N.A. Obtaining recombinant forms of the outer membrane protein F of Pseudomonas aeruginosa and assessment of their immunogenic properties. Appl. Biochem. Microbiol. 2011, 47 (8): 780-788.
  7. Sambrook J.F., Russell D.W. Molecular cloning, 2001.

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Copyright (c) 2012 Kaloshin A.A., Mikhaylova N.A., Leonova E.I.

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