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首页> 外文期刊>Infection and immunity >Identification of ragAB as a Temperature-Regulated Operon of Porphyromonas gingivalis W50 Using Differential Display of Randomly Primed RNA
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Identification of ragAB as a Temperature-Regulated Operon of Porphyromonas gingivalis W50 Using Differential Display of Randomly Primed RNA

机译：使用差异显示的随机引物RNA鉴定ragAB作为牙龈卟啉单胞菌W50的温度调节的操纵子。

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Porphyromonas gingivalis is a gram-negative, black-pigmented anaerobe that has been associated with advanced periodontal disease. The genome of P. gingivalis has the potential to produce a number of virulence determinants including proteases, hemagglutinins, hemolysin, invasion-associated proteins, and products of the pathogenicity island ragAB; however, little is known about how their expression is controlled. Periodontal pockets experience a higher temperature during inflammation, and this elevated temperature may influence the pathogenicity of P. gingivalis by changing its patterns of gene expression. In this study, RNA has been isolated from cells of P. gingivalisgrown to steady state at temperatures of 37, 39, and 41°C under hemin excess conditions (pH 7.0) in a chemostat. The RNA was subjected to PCR amplification following reverse transcription, using various combinations of randomly selected oligonucleotide primers. Reproducible RNA fingerprints have been obtained; however, differences were demonstrated in the RNA profiles of cells grown at the three temperatures, indicating differences in gene expression. Several PCR fragments were isolated that appeared to represent temperature-regulated genes. The nucleotide sequence of one of these has been identified as part of the ragAB locus, which codes for both a 55-kDa immunodominant antigen (RagB) and a homologue of the family of TonB-linked outer membrane receptors (RagA). These data indicate that expression of ragAB may be modulated in response to changes in temperature and that this may suggest a mechanism of evading the host response in the inflamed periodontal pocket.
机译： Porphyromonas gingivalis 是革兰氏阴性，黑色色素厌氧菌，与晚期牙周疾病有关。 P的基因组。牙龈炎具有产生多种毒性决定因素的潜力，包括蛋白酶，血凝素，溶血素，侵袭相关蛋白以及致病岛 ragAB 的产物。但是，关于如何控制它们的表达知之甚少。牙周袋在炎症过程中会经历较高的温度，而这种升高的温度可能会影响 P的致病性。改变牙龈的基因表达方式。在这项研究中，RNA已从 P细胞中分离出来。在恒化器中的血红素过量条件（pH 7.0）下，牙龈在37、39和41°C的温度下生长到稳定状态。在逆转录后，使用随机选择的寡核苷酸引物的各种组合对RNA进行PCR扩增。已获得可再现的RNA指纹图;但是，在这三个温度下生长的细胞的RNA图谱中显示出差异，表明基因表达的差异。分离了几个PCR片段，这些片段似乎代表了温度调节基因。其中之一的核苷酸序列已被鉴定为 ragAB 基因座的一部分，其编码55 kDa的免疫显性抗原（RagB）和与TonB连接的外膜受体家族的同系物。 （RagA）。这些数据表明， ragAB 的表达可以响应温度的变化而被调节，这可能暗示了在发炎的牙周袋中逃避宿主反应的机制。

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