Microarray-based identification of differentially expressed genes in intracellular Brucella abortus within RAW264.7 cells.

Brucella spp. is a species of facultative intracellular Gram-negative bacteria that induces abortion and causes sterility in domesticated mammals and chronic undulant fever in humans. Important determinants of Brucella's virulence and potential for chronic infection include the ability to circu...

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Main Authors: Mingxing Tian, Jing Qu, Xiangan Han, Min Zhang, Chan Ding, Jiabo Ding, Guanghua Chen, Shengqing Yu
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2013-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC3737221?pdf=render
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author Mingxing Tian
Jing Qu
Xiangan Han
Min Zhang
Chan Ding
Jiabo Ding
Guanghua Chen
Shengqing Yu
author_facet Mingxing Tian
Jing Qu
Xiangan Han
Min Zhang
Chan Ding
Jiabo Ding
Guanghua Chen
Shengqing Yu
author_sort Mingxing Tian
collection DOAJ
description Brucella spp. is a species of facultative intracellular Gram-negative bacteria that induces abortion and causes sterility in domesticated mammals and chronic undulant fever in humans. Important determinants of Brucella's virulence and potential for chronic infection include the ability to circumvent the host cell's internal surveillance system and the capability to proliferate within dedicated and non-dedicated phagocytes. Hence, identifying genes necessary for intracellular survival may hold the key to understanding Brucella infection. In the present study, microarray analysis reveals that 7.82% (244/3334) of all Brucella abortus genes were up-regulated and 5.4% (180/3334) were down-regulated in RAW264.7 cells, compared to free-living cells in TSB. qRT-PCR verification further confirmed a >5-fold up-regulation for fourteen genes. Functional analysis classified araC, ddp, and eryD as to partake in information storage and processing, alp, flgF and virB9 to be involved in cellular processes, hpcd and aldh to play a role in metabolism, mfs and nikC to be involved in both cellular processes and metabolism, and four hypothetical genes (bruAb1_1814, bruAb1_0475, bruAb1_1926, and bruAb1_0292) had unknown functions. Furthermore, we constructed a B. abortus 2308 mutant Δddp where the ddp gene is deleted in order to evaluate the role of ddp in intracellular survival. Infection assay indicated significantly higher adherence and invasion abilities of the Δddp mutant, however it does not survive well in RAW264.7 cells. Brucella may survive in hostile intracellular environment by modulating gene expression.
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spelling doaj.art-861535e30ebd42abbb3e3eabf5768eb62022-12-21T20:33:18ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0188e6701410.1371/journal.pone.0067014Microarray-based identification of differentially expressed genes in intracellular Brucella abortus within RAW264.7 cells.Mingxing TianJing QuXiangan HanMin ZhangChan DingJiabo DingGuanghua ChenShengqing YuBrucella spp. is a species of facultative intracellular Gram-negative bacteria that induces abortion and causes sterility in domesticated mammals and chronic undulant fever in humans. Important determinants of Brucella's virulence and potential for chronic infection include the ability to circumvent the host cell's internal surveillance system and the capability to proliferate within dedicated and non-dedicated phagocytes. Hence, identifying genes necessary for intracellular survival may hold the key to understanding Brucella infection. In the present study, microarray analysis reveals that 7.82% (244/3334) of all Brucella abortus genes were up-regulated and 5.4% (180/3334) were down-regulated in RAW264.7 cells, compared to free-living cells in TSB. qRT-PCR verification further confirmed a >5-fold up-regulation for fourteen genes. Functional analysis classified araC, ddp, and eryD as to partake in information storage and processing, alp, flgF and virB9 to be involved in cellular processes, hpcd and aldh to play a role in metabolism, mfs and nikC to be involved in both cellular processes and metabolism, and four hypothetical genes (bruAb1_1814, bruAb1_0475, bruAb1_1926, and bruAb1_0292) had unknown functions. Furthermore, we constructed a B. abortus 2308 mutant Δddp where the ddp gene is deleted in order to evaluate the role of ddp in intracellular survival. Infection assay indicated significantly higher adherence and invasion abilities of the Δddp mutant, however it does not survive well in RAW264.7 cells. Brucella may survive in hostile intracellular environment by modulating gene expression.http://europepmc.org/articles/PMC3737221?pdf=render
spellingShingle Mingxing Tian
Jing Qu
Xiangan Han
Min Zhang
Chan Ding
Jiabo Ding
Guanghua Chen
Shengqing Yu
Microarray-based identification of differentially expressed genes in intracellular Brucella abortus within RAW264.7 cells.
PLoS ONE
title Microarray-based identification of differentially expressed genes in intracellular Brucella abortus within RAW264.7 cells.
title_full Microarray-based identification of differentially expressed genes in intracellular Brucella abortus within RAW264.7 cells.
title_fullStr Microarray-based identification of differentially expressed genes in intracellular Brucella abortus within RAW264.7 cells.
title_full_unstemmed Microarray-based identification of differentially expressed genes in intracellular Brucella abortus within RAW264.7 cells.
title_short Microarray-based identification of differentially expressed genes in intracellular Brucella abortus within RAW264.7 cells.
title_sort microarray based identification of differentially expressed genes in intracellular brucella abortus within raw264 7 cells
url http://europepmc.org/articles/PMC3737221?pdf=render
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