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Determination of the expression of three fimbrial subunit proteins in cultured Trueperella pyogenes
BACKGROUND: Trueperella pyogenes is a commensal and a significant opportunistic pathogen in animals. A variety of identified or putative virulence factors are considered to significantly contribute to the occurrence of T. pyogenes infection in different species. However, these virulence factors are...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6134790/ https://www.ncbi.nlm.nih.gov/pubmed/30208923 http://dx.doi.org/10.1186/s13028-018-0407-3 |
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author | Liu, Mengcheng Wang, Bing Liang, Hongmin Ma, Bo Wang, Junwei Zhang, Wenlong |
author_facet | Liu, Mengcheng Wang, Bing Liang, Hongmin Ma, Bo Wang, Junwei Zhang, Wenlong |
author_sort | Liu, Mengcheng |
collection | PubMed |
description | BACKGROUND: Trueperella pyogenes is a commensal and a significant opportunistic pathogen in animals. A variety of identified or putative virulence factors are considered to significantly contribute to the occurrence of T. pyogenes infection in different species. However, these virulence factors are not fully understood. RESULTS: In the current study, the genes encoding putative fimbrial proteins, i.e. Fim A, Fim C, and Fim E, were cloned. Recombinant Fim A (rFim A), Fim C (rFim C), and Fim E (rFim E) were prepared and used to generate rabbit anti-rFim A, anti-rFim C, and anti-rFim E serum, respectively. Using these sera, we found that only Fim E was constitutively expressed in T. pyogenes. The expression level of Fim E in T. pyogenes peaked within 6–10 h of culture period in pH 7.5. Fim E protein expression was unaffected by anaerobic condition, but was inhibited by the microaerophilic condition. Tube agglutination tests indicated that Fim E was exhibited on the surface of T. pyogenes cells because anti-rFim E serum caused strong agglutination. Additionally, the blots for Fim A detection showed nonspecific reactions. Furthermore, the tube agglutination tests showed that anti-Fim A serum failed to cause agglutination of T. pyogenes cells, which indicated that Fim A was not, or poorly, expressed in cultured T. pyogenes. Anti-rFim C serum caused strong agglutination. However, the blots for Fim C detection showed a strong nonspecific reaction. Thus, the expression of Fim C was difficult to be determined using the current method. CONCLUSIONS: Fim E was expressed in cultured T. pyogenes. However, Fim A was either not or poorly expressed in cultured T. pyogenes. Moreover, Fim C expression was not determined using the current strategy. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13028-018-0407-3) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6134790 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-61347902018-09-15 Determination of the expression of three fimbrial subunit proteins in cultured Trueperella pyogenes Liu, Mengcheng Wang, Bing Liang, Hongmin Ma, Bo Wang, Junwei Zhang, Wenlong Acta Vet Scand Research BACKGROUND: Trueperella pyogenes is a commensal and a significant opportunistic pathogen in animals. A variety of identified or putative virulence factors are considered to significantly contribute to the occurrence of T. pyogenes infection in different species. However, these virulence factors are not fully understood. RESULTS: In the current study, the genes encoding putative fimbrial proteins, i.e. Fim A, Fim C, and Fim E, were cloned. Recombinant Fim A (rFim A), Fim C (rFim C), and Fim E (rFim E) were prepared and used to generate rabbit anti-rFim A, anti-rFim C, and anti-rFim E serum, respectively. Using these sera, we found that only Fim E was constitutively expressed in T. pyogenes. The expression level of Fim E in T. pyogenes peaked within 6–10 h of culture period in pH 7.5. Fim E protein expression was unaffected by anaerobic condition, but was inhibited by the microaerophilic condition. Tube agglutination tests indicated that Fim E was exhibited on the surface of T. pyogenes cells because anti-rFim E serum caused strong agglutination. Additionally, the blots for Fim A detection showed nonspecific reactions. Furthermore, the tube agglutination tests showed that anti-Fim A serum failed to cause agglutination of T. pyogenes cells, which indicated that Fim A was not, or poorly, expressed in cultured T. pyogenes. Anti-rFim C serum caused strong agglutination. However, the blots for Fim C detection showed a strong nonspecific reaction. Thus, the expression of Fim C was difficult to be determined using the current method. CONCLUSIONS: Fim E was expressed in cultured T. pyogenes. However, Fim A was either not or poorly expressed in cultured T. pyogenes. Moreover, Fim C expression was not determined using the current strategy. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13028-018-0407-3) contains supplementary material, which is available to authorized users. BioMed Central 2018-09-12 /pmc/articles/PMC6134790/ /pubmed/30208923 http://dx.doi.org/10.1186/s13028-018-0407-3 Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Liu, Mengcheng Wang, Bing Liang, Hongmin Ma, Bo Wang, Junwei Zhang, Wenlong Determination of the expression of three fimbrial subunit proteins in cultured Trueperella pyogenes |
title | Determination of the expression of three fimbrial subunit proteins in cultured Trueperella pyogenes |
title_full | Determination of the expression of three fimbrial subunit proteins in cultured Trueperella pyogenes |
title_fullStr | Determination of the expression of three fimbrial subunit proteins in cultured Trueperella pyogenes |
title_full_unstemmed | Determination of the expression of three fimbrial subunit proteins in cultured Trueperella pyogenes |
title_short | Determination of the expression of three fimbrial subunit proteins in cultured Trueperella pyogenes |
title_sort | determination of the expression of three fimbrial subunit proteins in cultured trueperella pyogenes |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6134790/ https://www.ncbi.nlm.nih.gov/pubmed/30208923 http://dx.doi.org/10.1186/s13028-018-0407-3 |
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