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Investigation of TbfA in Riemerella anatipestifer using plasmid-based methods for gene over-expression and knockdown
Riemerella anatipestifer is a duck pathogen that has caused serious economic losses to the duck industry worldwide. Despite this, there are few reported studies of the physiological and pathogenic mechanisms of Riemerella anatipestifer infection. In previous study, we have shown that TonB1 and TonB2...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5109031/ https://www.ncbi.nlm.nih.gov/pubmed/27845444 http://dx.doi.org/10.1038/srep37159 |
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author | Liu, MaFeng Wang, MengYi Zhu, DeKang Wang, MingShu Jia, RenYong Chen, Shun Sun, KunFeng Yang, Qiao Wu, Ying Chen, XiaoYue Biville, Francis Cheng, AnChun |
author_facet | Liu, MaFeng Wang, MengYi Zhu, DeKang Wang, MingShu Jia, RenYong Chen, Shun Sun, KunFeng Yang, Qiao Wu, Ying Chen, XiaoYue Biville, Francis Cheng, AnChun |
author_sort | Liu, MaFeng |
collection | PubMed |
description | Riemerella anatipestifer is a duck pathogen that has caused serious economic losses to the duck industry worldwide. Despite this, there are few reported studies of the physiological and pathogenic mechanisms of Riemerella anatipestifer infection. In previous study, we have shown that TonB1 and TonB2 were involved in hemin uptake. TonB family protein (TbfA) was not investigated, since knockout of this gene was not successful at that time. Here, we used a plasmid based gene over-expression and knockdown to investigate its function. First, we constructed three Escherichia-Riemerella anatipestifer shuttle vectors containing three different native Riemerella anatipestifer promoters. The shuttle plasmids were introduced into Riemerella anatipestifer ATCC11845 by conjugation at an efficiency of 5 × 10(−5) antibiotic-resistant transconjugants per recipient cell. Based on the high-expression shuttle vector pLMF03, a method for gene knockdown was established. Knockdown of TbfA in Riemerella anatipestifer ATCC11845 decreased the organism’s growth ability in TSB medium but did not affect its hemin utilization. In contrast, over-expression of TbfA in Riemerella anatipestifer ATCC11845ΔtonB1ΔtonB2. Significantly promoted the organism’s growth in TSB medium but significantly inhibited its hemin utilization. Collectively, these findings suggest that TbfA is not involved in hemin utilization by Riemerella anatipestifer. |
format | Online Article Text |
id | pubmed-5109031 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-51090312016-11-25 Investigation of TbfA in Riemerella anatipestifer using plasmid-based methods for gene over-expression and knockdown Liu, MaFeng Wang, MengYi Zhu, DeKang Wang, MingShu Jia, RenYong Chen, Shun Sun, KunFeng Yang, Qiao Wu, Ying Chen, XiaoYue Biville, Francis Cheng, AnChun Sci Rep Article Riemerella anatipestifer is a duck pathogen that has caused serious economic losses to the duck industry worldwide. Despite this, there are few reported studies of the physiological and pathogenic mechanisms of Riemerella anatipestifer infection. In previous study, we have shown that TonB1 and TonB2 were involved in hemin uptake. TonB family protein (TbfA) was not investigated, since knockout of this gene was not successful at that time. Here, we used a plasmid based gene over-expression and knockdown to investigate its function. First, we constructed three Escherichia-Riemerella anatipestifer shuttle vectors containing three different native Riemerella anatipestifer promoters. The shuttle plasmids were introduced into Riemerella anatipestifer ATCC11845 by conjugation at an efficiency of 5 × 10(−5) antibiotic-resistant transconjugants per recipient cell. Based on the high-expression shuttle vector pLMF03, a method for gene knockdown was established. Knockdown of TbfA in Riemerella anatipestifer ATCC11845 decreased the organism’s growth ability in TSB medium but did not affect its hemin utilization. In contrast, over-expression of TbfA in Riemerella anatipestifer ATCC11845ΔtonB1ΔtonB2. Significantly promoted the organism’s growth in TSB medium but significantly inhibited its hemin utilization. Collectively, these findings suggest that TbfA is not involved in hemin utilization by Riemerella anatipestifer. Nature Publishing Group 2016-11-15 /pmc/articles/PMC5109031/ /pubmed/27845444 http://dx.doi.org/10.1038/srep37159 Text en Copyright © 2016, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Liu, MaFeng Wang, MengYi Zhu, DeKang Wang, MingShu Jia, RenYong Chen, Shun Sun, KunFeng Yang, Qiao Wu, Ying Chen, XiaoYue Biville, Francis Cheng, AnChun Investigation of TbfA in Riemerella anatipestifer using plasmid-based methods for gene over-expression and knockdown |
title | Investigation of TbfA in Riemerella anatipestifer using plasmid-based methods for gene over-expression and knockdown |
title_full | Investigation of TbfA in Riemerella anatipestifer using plasmid-based methods for gene over-expression and knockdown |
title_fullStr | Investigation of TbfA in Riemerella anatipestifer using plasmid-based methods for gene over-expression and knockdown |
title_full_unstemmed | Investigation of TbfA in Riemerella anatipestifer using plasmid-based methods for gene over-expression and knockdown |
title_short | Investigation of TbfA in Riemerella anatipestifer using plasmid-based methods for gene over-expression and knockdown |
title_sort | investigation of tbfa in riemerella anatipestifer using plasmid-based methods for gene over-expression and knockdown |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5109031/ https://www.ncbi.nlm.nih.gov/pubmed/27845444 http://dx.doi.org/10.1038/srep37159 |
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