Ectopic Expression of O Antigen in <named-content content-type="genus-species">Bordetella pertussis</named-content> by a Novel Genomic Integration System

ABSTRACT We describe a novel genome integration system that enables the introduction of DNA fragments as large as 50 kbp into the chromosomes of recipient bacteria. This system, named BPI, comprises a bacterial artificial chromosome vector and phage-derived gene integration machinery. We introduced...

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Autores principales:	Keisuke Ishigaki, Naoaki Shinzawa, Sayaka Nishikawa, Koichiro Suzuki, Aya Fukui-Miyazaki, Yasuhiko Horiguchi
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	Bordetella Bordetella bronchiseptica Bordetella pertussis O antigen site-specific recombination Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/ccb87c7d76c8458daaff9033d6fb1c04
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spelling	oai:doaj.org-article:ccb87c7d76c8458daaff9033d6fb1c042021-11-15T15:22:01ZEctopic Expression of O Antigen in <named-content content-type="genus-species">Bordetella pertussis</named-content> by a Novel Genomic Integration System10.1128/mSphere.00417-172379-5042https://doaj.org/article/ccb87c7d76c8458daaff9033d6fb1c042018-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00417-17https://doaj.org/toc/2379-5042ABSTRACT We describe a novel genome integration system that enables the introduction of DNA fragments as large as 50 kbp into the chromosomes of recipient bacteria. This system, named BPI, comprises a bacterial artificial chromosome vector and phage-derived gene integration machinery. We introduced the wbm locus of Bordetella bronchiseptica, which is required for O antigen biosynthesis, into the chromosome of B. pertussis, which intrinsically lacks O antigen, using the BPI system. After the introduction of the wbm locus, B. pertussis presented an additional substance in the lipooligosaccharide fraction that was specifically recognized by the anti-B. bronchiseptica antibody but not the anti-B. pertussis antibody, indicating that B. pertussis expressed O antigen corresponding to that of B. bronchiseptica. O antigen-expressing B. pertussis was less sensitive to the bactericidal effects of serum and polymyxin B than the isogenic parental strain. In addition, an in vivo competitive infection assay showed that O antigen-expressing B. pertussis dominantly colonized the mouse respiratory tract over the parental strain. These results indicate that the BPI system provides a means to alter the phenotypes of bacteria by introducing large exogenous DNA fragments. IMPORTANCE Some bacterial phenotypes emerge through the cooperative functions of a number of genes residing within a large genetic locus. To transfer the phenotype of one bacterium to another, a means to introduce the large genetic locus into the recipient bacterium is needed. Therefore, we developed a novel system by combining the advantages of a bacterial artificial chromosome vector and phage-derived gene integration machinery. In this study, we succeeded for the first time in introducing a gene locus involved in O antigen biosynthesis of Bordetella bronchiseptica into the chromosome of B. pertussis, which intrinsically lacks O antigen, and using this system we analyzed phenotypic alterations in the resultant mutant strain of B. pertussis. The present results demonstrate that this system successfully accomplished the above-described purpose. We consider this system to be applicable to a number of bacteria other than Bordetella.Keisuke IshigakiNaoaki ShinzawaSayaka NishikawaKoichiro SuzukiAya Fukui-MiyazakiYasuhiko HoriguchiAmerican Society for MicrobiologyarticleBordetellaBordetella bronchisepticaBordetella pertussisO antigensite-specific recombinationMicrobiologyQR1-502ENmSphere, Vol 3, Iss 1 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Bordetella Bordetella bronchiseptica Bordetella pertussis O antigen site-specific recombination Microbiology QR1-502
spellingShingle	Bordetella Bordetella bronchiseptica Bordetella pertussis O antigen site-specific recombination Microbiology QR1-502 Keisuke Ishigaki Naoaki Shinzawa Sayaka Nishikawa Koichiro Suzuki Aya Fukui-Miyazaki Yasuhiko Horiguchi Ectopic Expression of O Antigen in <named-content content-type="genus-species">Bordetella pertussis</named-content> by a Novel Genomic Integration System
description	ABSTRACT We describe a novel genome integration system that enables the introduction of DNA fragments as large as 50 kbp into the chromosomes of recipient bacteria. This system, named BPI, comprises a bacterial artificial chromosome vector and phage-derived gene integration machinery. We introduced the wbm locus of Bordetella bronchiseptica, which is required for O antigen biosynthesis, into the chromosome of B. pertussis, which intrinsically lacks O antigen, using the BPI system. After the introduction of the wbm locus, B. pertussis presented an additional substance in the lipooligosaccharide fraction that was specifically recognized by the anti-B. bronchiseptica antibody but not the anti-B. pertussis antibody, indicating that B. pertussis expressed O antigen corresponding to that of B. bronchiseptica. O antigen-expressing B. pertussis was less sensitive to the bactericidal effects of serum and polymyxin B than the isogenic parental strain. In addition, an in vivo competitive infection assay showed that O antigen-expressing B. pertussis dominantly colonized the mouse respiratory tract over the parental strain. These results indicate that the BPI system provides a means to alter the phenotypes of bacteria by introducing large exogenous DNA fragments. IMPORTANCE Some bacterial phenotypes emerge through the cooperative functions of a number of genes residing within a large genetic locus. To transfer the phenotype of one bacterium to another, a means to introduce the large genetic locus into the recipient bacterium is needed. Therefore, we developed a novel system by combining the advantages of a bacterial artificial chromosome vector and phage-derived gene integration machinery. In this study, we succeeded for the first time in introducing a gene locus involved in O antigen biosynthesis of Bordetella bronchiseptica into the chromosome of B. pertussis, which intrinsically lacks O antigen, and using this system we analyzed phenotypic alterations in the resultant mutant strain of B. pertussis. The present results demonstrate that this system successfully accomplished the above-described purpose. We consider this system to be applicable to a number of bacteria other than Bordetella.
format	article
author	Keisuke Ishigaki Naoaki Shinzawa Sayaka Nishikawa Koichiro Suzuki Aya Fukui-Miyazaki Yasuhiko Horiguchi
author_facet	Keisuke Ishigaki Naoaki Shinzawa Sayaka Nishikawa Koichiro Suzuki Aya Fukui-Miyazaki Yasuhiko Horiguchi
author_sort	Keisuke Ishigaki
title	Ectopic Expression of O Antigen in <named-content content-type="genus-species">Bordetella pertussis</named-content> by a Novel Genomic Integration System
title_short	Ectopic Expression of O Antigen in <named-content content-type="genus-species">Bordetella pertussis</named-content> by a Novel Genomic Integration System
title_full	Ectopic Expression of O Antigen in <named-content content-type="genus-species">Bordetella pertussis</named-content> by a Novel Genomic Integration System
title_fullStr	Ectopic Expression of O Antigen in <named-content content-type="genus-species">Bordetella pertussis</named-content> by a Novel Genomic Integration System
title_full_unstemmed	Ectopic Expression of O Antigen in <named-content content-type="genus-species">Bordetella pertussis</named-content> by a Novel Genomic Integration System
title_sort	ectopic expression of o antigen in <named-content content-type="genus-species">bordetella pertussis</named-content> by a novel genomic integration system
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/ccb87c7d76c8458daaff9033d6fb1c04
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Ectopic Expression of O Antigen in <named-content content-type="genus-species">Bordetella pertussis</named-content> by a Novel Genomic Integration System

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