Transcriptional Analysis of Temporal Gene Expression in Germinating Clostridium difficile 630 Endospores
Transcriptional Analysis of Temporal Gene Expression in Germinating Clostridium difficile 630 Endospores
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DOI：
10.1371/journal.pone.0064011
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发表时间：
2013-05-15
期刊：
影响因子：
3.7
通讯作者：
Fairweather, Neil F.
中科院分区：
文献类型：
--
作者：
Dembek, Marcin;Stabler, Richard A.;Fairweather, Neil F.
艰难梭菌是工业化国家医院获得性腹泻的主要原因。在不利于生长的条件下，病原体通过不对称细胞分裂产生代谢休眠的内生孢子。它们对化学和物理压力都具有极强的抵抗力，并提供了艰难梭菌可以逃避暴露于热、氧气、酒精和某些消毒剂的潜在致命后果的机制。孢子是主要的感染因子，必须发芽才能允许营养细胞生长和产生毒素。虽然芽孢杆菌中的孢子萌发已被充分了解，但对于艰难梭菌的萌发和生长却知之甚少。在这里，我们使用全基因组转录分析来阐明艰难梭菌 630 内生孢子萌发中的时间基因表达模式。我们拥有大规模生产和纯化孢子的优化方法。纯化孢子的萌发特性已得到表征，RNA 提取方案也已得到优化。基因表达在发芽和生长过程中是高度动态的，并且被发现涉及大量基因。利用这种全基因组微阵列方法，我们鉴定了 511 个在艰难梭菌萌发过程中显着上调或下调的基因 (p
Clostridium difficile is the leading cause of hospital acquired diarrhoea in industrialised countries. Under conditions that are not favourable for growth, the pathogen produces metabolically dormant endospores via asymmetric cell division. These are extremely resistant to both chemical and physical stress and provide the mechanism by which C. difficile can evade the potentially fatal consequences of exposure to heat, oxygen, alcohol, and certain disinfectants. Spores are the primary infective agent and must germinate to allow for vegetative cell growth and toxin production. While spore germination in Bacillus is well understood, little is known about C. difficile germination and outgrowth. Here we use genome-wide transcriptional analysis to elucidate the temporal gene expression patterns in C. difficile 630 endospore germination. We have optimized methods for large scale production and purification of spores. The germination characteristics of purified spores have been characterized and RNA extraction protocols have been optimized. Gene expression was highly dynamic during germination and outgrowth, and was found to involve a large number of genes. Using this genome-wide, microarray approach we have identified 511 genes that are significantly up-or down-regulated during C. difficile germination (p