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首页> 外文期刊>Microbial Pathogenesis >GAPDH, rhbC, and vapA gene expression in Rhodococcus equi cultured under different iron concentrations
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GAPDH, rhbC, and vapA gene expression in Rhodococcus equi cultured under different iron concentrations

机译：在不同铁浓度下越rhodococcus equi中的Gapdh，RHBC和VAPA基因表达

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The ability of Rhodococcus equi to survive in macrophages and cause pneumonia in foals depends on vapA and rhbC genes, which produce the virulence-associated protein A (VapA) and the rhequichelin siderophore, respectively. Virulent R. equi acquires Fe from transferrin by unknown mechanisms. Our objectives were to determine the role of GAPDH in Fe homeostasis, to further characterize GAPDH, rhbC, and vapA expression under iron homeostasis, and to document the occurrence of rhbC gene in R. equi isolates. Therefore, vapA + R. equi was cultured under excessive, physiologic, and restricted iron concentrations, and quantitative culture and gene expression were performed. The relative expression of GAPDH, rhbC, and vapA after 48 h of culture were analyzed by qPCR. To determine the rhbC occurrence, total DNA was extracted from R. equi isolated from foals with clinical rhodococcosis (n = 22), healthy horses (feces, n = 16; nasal swab, n = 9), soil (n = 6), and 2 ATCC reference strains. Conventional PCR was performed to identify genus/species, vapA, and rhbC genes. Iron restriction proportionally decreased R. equi growth rates, and induced high expression of both GAPDH and vapA. The putative role of GAPDH in R. equi iron homeostasis should be further investigated. rhbC was significantly up-regulated under both Fe excess and critical starvation. The rhbC gene was identified in all clinical isolates and soil, but it was absent in 2 isolates from healthy horses, suggesting that rhequichelin is not required for R. equi nasal and intestinal colonization.
机译：rhodococcus在巨噬细胞中生存的能力并导致肺炎的肺炎依赖于VAPA和RHBC基因，分别产生毒力相关的蛋白A（VAPA）和rhequichelin Siderophore。毒性R. Equi通过未知机制从转铁蛋白获取Fe。我们的目标是确定GAPDH在Fe HoosoTasis的作用，进一步表征铁稳态下的GAPDH，RHBC和VAPA表达，并记录R. Equi分离株中的RHBC基因的发生。因此，在过量，生理学和限制的铁浓度下培养VAPA + R.培养，并进行定量培养和基因表达。通过QPCR分析GAPDH，RHBC和VAPA后的相对表达。为了确定RHBC的发生，从临床核疾病（n = 22），健康马（粪便，n = 16;鼻拭子，n = 9），土壤（n = 6），从R. Qual中提取总DNA。和2个ATCC参考菌株。进行常规PCR以鉴定属/物种，VAPA和RHBC基因。铁限制比例地降低了R. Equi生长率，并诱导了GAPDH和VAPA的高表达。 GAPDH在R. Equi Iron Sopostasis的推定作用应进一步调查。在Fe超额和临界饥饿下，RHBC显着上调。在所有临床分离物和土壤中鉴定了rHBC基因，但它不存在于2个来自健康马的分离物中，这表明R.等鼻和肠道殖民化不需要rhequichelin。

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