Cargando…

Molecular study on the carAB operon reveals that carB gene is required for swimming and biofilm formation in Xanthomonas citri subsp. citri

BACKGROUND: The carA and carB genes code the small and large subunits of carbamoyl-phosphate synthase (CPS) that responsible for arginine and pyrimidine production. The purpose of this work was to study the gene organization and expression pattern of carAB operon, and the biological functions of car...

Descripción completa

Detalles Bibliográficos
Autores principales: Zhuo, Tao, Rou, Wei, Song, Xue, Guo, Jing, Fan, Xiaojing, Kamau, Gicharu Gibson, Zou, Huasong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4619228/
https://www.ncbi.nlm.nih.gov/pubmed/26494007
http://dx.doi.org/10.1186/s12866-015-0555-9
_version_ 1782397053939220480
author Zhuo, Tao
Rou, Wei
Song, Xue
Guo, Jing
Fan, Xiaojing
Kamau, Gicharu Gibson
Zou, Huasong
author_facet Zhuo, Tao
Rou, Wei
Song, Xue
Guo, Jing
Fan, Xiaojing
Kamau, Gicharu Gibson
Zou, Huasong
author_sort Zhuo, Tao
collection PubMed
description BACKGROUND: The carA and carB genes code the small and large subunits of carbamoyl-phosphate synthase (CPS) that responsible for arginine and pyrimidine production. The purpose of this work was to study the gene organization and expression pattern of carAB operon, and the biological functions of carA and carB genes in Xanthomonas citri subsp. citri. METHODS: RT-PCR method was employed to identify the full length of carAB operon transcript in X. citri subsp. citri. The promoter of carAB operon was predicted and analyzed its activity by fusing a GUS reporter gene. The swimming motility was tested on 0.25 % agar NY plates with 1 % glucose. Biofilm was measured by cell adhesion to polyvinyl chloride 96-well plate. RESULTS: The results indicated that carAB operon was composed of five gene members carA-orf-carB-greA-rpfE. A single promoter was predicted from the nucleotide sequence upstream of carAB operon, and its sensitivity to glutamic acid, uracil and arginine was confirmed by fusing a GUS reporter gene. Deletion mutagenesis of carB gene resulted in reduced abilities in swimming on soft solid media and in forming biofilm on polystyrene microtiter plates. CONCLUSIONS: From these results, we concluded that carAB operon was involved in multiple biological processes in X. citri subsp. citri. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12866-015-0555-9) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-4619228
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-46192282015-10-26 Molecular study on the carAB operon reveals that carB gene is required for swimming and biofilm formation in Xanthomonas citri subsp. citri Zhuo, Tao Rou, Wei Song, Xue Guo, Jing Fan, Xiaojing Kamau, Gicharu Gibson Zou, Huasong BMC Microbiol Research Article BACKGROUND: The carA and carB genes code the small and large subunits of carbamoyl-phosphate synthase (CPS) that responsible for arginine and pyrimidine production. The purpose of this work was to study the gene organization and expression pattern of carAB operon, and the biological functions of carA and carB genes in Xanthomonas citri subsp. citri. METHODS: RT-PCR method was employed to identify the full length of carAB operon transcript in X. citri subsp. citri. The promoter of carAB operon was predicted and analyzed its activity by fusing a GUS reporter gene. The swimming motility was tested on 0.25 % agar NY plates with 1 % glucose. Biofilm was measured by cell adhesion to polyvinyl chloride 96-well plate. RESULTS: The results indicated that carAB operon was composed of five gene members carA-orf-carB-greA-rpfE. A single promoter was predicted from the nucleotide sequence upstream of carAB operon, and its sensitivity to glutamic acid, uracil and arginine was confirmed by fusing a GUS reporter gene. Deletion mutagenesis of carB gene resulted in reduced abilities in swimming on soft solid media and in forming biofilm on polystyrene microtiter plates. CONCLUSIONS: From these results, we concluded that carAB operon was involved in multiple biological processes in X. citri subsp. citri. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12866-015-0555-9) contains supplementary material, which is available to authorized users. BioMed Central 2015-10-23 /pmc/articles/PMC4619228/ /pubmed/26494007 http://dx.doi.org/10.1186/s12866-015-0555-9 Text en © Zhuo et al. 2015 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 Article
Zhuo, Tao
Rou, Wei
Song, Xue
Guo, Jing
Fan, Xiaojing
Kamau, Gicharu Gibson
Zou, Huasong
Molecular study on the carAB operon reveals that carB gene is required for swimming and biofilm formation in Xanthomonas citri subsp. citri
title Molecular study on the carAB operon reveals that carB gene is required for swimming and biofilm formation in Xanthomonas citri subsp. citri
title_full Molecular study on the carAB operon reveals that carB gene is required for swimming and biofilm formation in Xanthomonas citri subsp. citri
title_fullStr Molecular study on the carAB operon reveals that carB gene is required for swimming and biofilm formation in Xanthomonas citri subsp. citri
title_full_unstemmed Molecular study on the carAB operon reveals that carB gene is required for swimming and biofilm formation in Xanthomonas citri subsp. citri
title_short Molecular study on the carAB operon reveals that carB gene is required for swimming and biofilm formation in Xanthomonas citri subsp. citri
title_sort molecular study on the carab operon reveals that carb gene is required for swimming and biofilm formation in xanthomonas citri subsp. citri
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4619228/
https://www.ncbi.nlm.nih.gov/pubmed/26494007
http://dx.doi.org/10.1186/s12866-015-0555-9
work_keys_str_mv AT zhuotao molecularstudyonthecaraboperonrevealsthatcarbgeneisrequiredforswimmingandbiofilmformationinxanthomonascitrisubspcitri
AT rouwei molecularstudyonthecaraboperonrevealsthatcarbgeneisrequiredforswimmingandbiofilmformationinxanthomonascitrisubspcitri
AT songxue molecularstudyonthecaraboperonrevealsthatcarbgeneisrequiredforswimmingandbiofilmformationinxanthomonascitrisubspcitri
AT guojing molecularstudyonthecaraboperonrevealsthatcarbgeneisrequiredforswimmingandbiofilmformationinxanthomonascitrisubspcitri
AT fanxiaojing molecularstudyonthecaraboperonrevealsthatcarbgeneisrequiredforswimmingandbiofilmformationinxanthomonascitrisubspcitri
AT kamaugicharugibson molecularstudyonthecaraboperonrevealsthatcarbgeneisrequiredforswimmingandbiofilmformationinxanthomonascitrisubspcitri
AT zouhuasong molecularstudyonthecaraboperonrevealsthatcarbgeneisrequiredforswimmingandbiofilmformationinxanthomonascitrisubspcitri