CONSTRUCTION OF RECOMBINANT ATTENUATED BORDETELLA PERTUSSIS BACTERIA OF PTXP3 GENOTYPE
Aim. The construction of recombinant attenuated Bordetella pertussis bacteria of ptxP3 genotype and its genetic and biological stability characteristics. Materials and methods. During construction of recombinant attenuated bacteria of ptxP3 genotype virulent Bordetella pertussis bacteria of ptxP1 ge...
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Central Research Institute for Epidemiology
2018-08-01
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doaj-cf5bde6925b849c79904007b1955626a2021-07-28T13:07:51ZrusCentral Research Institute for Epidemiology Журнал микробиологии, эпидемиологии и иммунобиологии0372-93112686-76132018-08-0104334110.36233/0372-9311-2018-4-33-41284CONSTRUCTION OF RECOMBINANT ATTENUATED BORDETELLA PERTUSSIS BACTERIA OF PTXP3 GENOTYPEE. G. Semin0L. N. Sinyashina1A. Yu. Medkova2G. I. Karataev3Gamaleya National Research Centre of Epidemiology and MicrobiologyGamaleya National Research Centre of Epidemiology and MicrobiologyGamaleya National Research Centre of Epidemiology and MicrobiologyGamaleya National Research Centre of Epidemiology and MicrobiologyAim. The construction of recombinant attenuated Bordetella pertussis bacteria of ptxP3 genotype and its genetic and biological stability characteristics. Materials and methods. During construction of recombinant attenuated bacteria of ptxP3 genotype virulent Bordetella pertussis bacteria of ptxP1 genotype were used as a recipient. PtxP1 genotype bacteria are used for whole cell pertussis (wP) vaccine production in Russia. Mutant bacteria B. pertussis 475 were received by crossing-over between chromosome comprising native copy of target sequence and its mutant copy in recombinant suicide plasmid transferred in recipient bacteria by conjugation. Genetically engineered construction of recombinant plasmids was conducted. The structure of modified chromosome locus of attenuated bacteria was determined by PCR and amplification fragments sequence. The stability of structure and characteristics of attenuated bacteria was defined after 15 passages of strains on culture medium and 5 passages in mice. Results. Isogenic attenuated ptxP1 B. pertussis 4M and ptxP3 B. pertussis 4MKS were constructed. These bacteria produce non-toxic pertussis toxin (PT) and do not produce dermonecrotic toxin (DNT). The promoter region of ptx operon contains mutation, typical for «new» genotype of circulating virulent bacteria and increasing PT production. The structure of modified DNA fragments and characteristics of attenuated bacteria did not change while storing and after passages on culture medium and in mice. Conclusion. Recombinant attenuated bacteria B. pertussis 4MKS of «new» ptxP3 genotype are constructed. Application perspectiveness of genetically engineered modification of isogenic B.pertussis bacteria for pertussis vaccines development is shown.https://microbiol.elpub.ru/jour/article/view/284pertussisrecombinant attenuated bacteria b. pertussisconstructiongenotypevaccines |
collection |
DOAJ |
language |
Russian |
format |
Article |
sources |
DOAJ |
author |
E. G. Semin L. N. Sinyashina A. Yu. Medkova G. I. Karataev |
spellingShingle |
E. G. Semin L. N. Sinyashina A. Yu. Medkova G. I. Karataev CONSTRUCTION OF RECOMBINANT ATTENUATED BORDETELLA PERTUSSIS BACTERIA OF PTXP3 GENOTYPE Журнал микробиологии, эпидемиологии и иммунобиологии pertussis recombinant attenuated bacteria b. pertussis construction genotype vaccines |
author_facet |
E. G. Semin L. N. Sinyashina A. Yu. Medkova G. I. Karataev |
author_sort |
E. G. Semin |
title |
CONSTRUCTION OF RECOMBINANT ATTENUATED BORDETELLA PERTUSSIS BACTERIA OF PTXP3 GENOTYPE |
title_short |
CONSTRUCTION OF RECOMBINANT ATTENUATED BORDETELLA PERTUSSIS BACTERIA OF PTXP3 GENOTYPE |
title_full |
CONSTRUCTION OF RECOMBINANT ATTENUATED BORDETELLA PERTUSSIS BACTERIA OF PTXP3 GENOTYPE |
title_fullStr |
CONSTRUCTION OF RECOMBINANT ATTENUATED BORDETELLA PERTUSSIS BACTERIA OF PTXP3 GENOTYPE |
title_full_unstemmed |
CONSTRUCTION OF RECOMBINANT ATTENUATED BORDETELLA PERTUSSIS BACTERIA OF PTXP3 GENOTYPE |
title_sort |
construction of recombinant attenuated bordetella pertussis bacteria of ptxp3 genotype |
publisher |
Central Research Institute for Epidemiology |
series |
Журнал микробиологии, эпидемиологии и иммунобиологии |
issn |
0372-9311 2686-7613 |
publishDate |
2018-08-01 |
description |
Aim. The construction of recombinant attenuated Bordetella pertussis bacteria of ptxP3 genotype and its genetic and biological stability characteristics. Materials and methods. During construction of recombinant attenuated bacteria of ptxP3 genotype virulent Bordetella pertussis bacteria of ptxP1 genotype were used as a recipient. PtxP1 genotype bacteria are used for whole cell pertussis (wP) vaccine production in Russia. Mutant bacteria B. pertussis 475 were received by crossing-over between chromosome comprising native copy of target sequence and its mutant copy in recombinant suicide plasmid transferred in recipient bacteria by conjugation. Genetically engineered construction of recombinant plasmids was conducted. The structure of modified chromosome locus of attenuated bacteria was determined by PCR and amplification fragments sequence. The stability of structure and characteristics of attenuated bacteria was defined after 15 passages of strains on culture medium and 5 passages in mice. Results. Isogenic attenuated ptxP1 B. pertussis 4M and ptxP3 B. pertussis 4MKS were constructed. These bacteria produce non-toxic pertussis toxin (PT) and do not produce dermonecrotic toxin (DNT). The promoter region of ptx operon contains mutation, typical for «new» genotype of circulating virulent bacteria and increasing PT production. The structure of modified DNA fragments and characteristics of attenuated bacteria did not change while storing and after passages on culture medium and in mice. Conclusion. Recombinant attenuated bacteria B. pertussis 4MKS of «new» ptxP3 genotype are constructed. Application perspectiveness of genetically engineered modification of isogenic B.pertussis bacteria for pertussis vaccines development is shown. |
topic |
pertussis recombinant attenuated bacteria b. pertussis construction genotype vaccines |
url |
https://microbiol.elpub.ru/jour/article/view/284 |
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