Expression and regulation of the ery operon of Brucella melitensis in human trophoblast cells
Expression and regulation of the ery operon of Brucella melitensis in human trophoblast cells
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羊种布鲁氏菌ery操纵子在人滋养层细胞中的表达及调控
DOI：
10.3892/etm.2016.3688
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发表时间：
2016-10-01
影响因子：
2.7
通讯作者：
Chen, Chuangfu
中科院分区：
文献类型：
--
作者：
Zhang, Hui;Dou, Xiaoxia;Chen, Chuangfu
布鲁氏菌病主要是家畜的一种疾病，其中细菌定位于胎儿组织，如胚胎滋养层细胞和含有刺激布鲁氏菌属的细菌的液体。增长布鲁氏菌属的一个特点是对细菌的利用。ery操纵子包含4个基因（eryA、eryB、eryC和eryD），用于利用细菌的能量，并在布鲁氏菌的生存和繁殖中起主要作用。本研究的目的是对布鲁氏菌感染胚胎滋养层细胞（HPT-8细胞）后不同时间点ery操纵子基因表达的差异进行初步表征。结果表明，ery操纵子在HPT-8细胞中的表达量高于培养液。在HPT-8细胞中，eryA、eryB和eryC的相对表达在感染后2 h达到峰值，而eryD的表达在感染后3 h达到峰值。增加的eryD表达可抑制eryA、eryB和eryC的表达。然而，ery操纵子的表达是稳定的，在细胞中的存在下。通过同源重组成功构建了ery操纵子的2308 ery和027 ery突变体，并在RAW 264.7小鼠巨噬细胞中进行了减毒。ery操纵子基因的特性及其在布鲁氏菌感染中的表达谱进一步有助于我们理解布鲁氏菌病感染的分子机制和发病机制。
Brucellosis is primarily a disease of domestic animals in which the bacteria localizes to fetal tissues such as embryonic trophoblast cells and fluids containing erythritol, which stimulates Brucella spp. growth. The utilization of erythritol is a characteristic of the genus Brucella. The ery operon contains four genes (eryA, eryB, eryC and eryD) for the utilization of erythritol, and plays a major role in the survival and multiplication of Brucella spp. The objective of the present study was to conduct a preliminary characterization of differential genes expression of the ery operon at several time points after Brucella infected embryonic trophoblast cells (HPT-8 cells). The result showed that the ery operon expression was higher in HPT-8 cells compared with the medium. The relative expression of eryA, eryB and eryC peaked at 2 h post-infection in HPT-8 cells, and eryD expression peaked at 3 h post-infection. The expression of eryA, eryB and eryC may be inhibited by increased eryD expression. However, the expression of the ery operon was stable in the presence of erythritol in cells. 2308ery and 027ery mutants of the ery operon were successfully constructed by homologous recombination, which were attenuated in RAW 264.7 murine macrophages. The characterization of the ery operon genes and their expression profiles in response to Brucella infection further contributes to our understanding of the molecular mechanisms of infection and the pathogenesis of brucellosis.