Restriction-modification mediated barriers to exogenous DNA uptake and incorporation employed by Prevotella intermedia.
Restriction-modification mediated barriers to exogenous DNA uptake and incorporation employed by Prevotella intermedia.
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DOI：
10.1371/journal.pone.0185234
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Rittling SR
中科院分区：
文献类型：
--
作者：
Johnston CD;Skeete CA;Fomenkov A;Roberts RJ;Rittling SR
中间普雷沃菌是一种重要的牙周致病菌，与人类呼吸道和囊性纤维化肺部感染的关系日益密切。然而，这种病原体所采用的具体机制仍然只有部分特征和知之甚少，主要是由于其完全缺乏遗传可及性。在这里，除了克隆和限制性分析之外，我们还使用单分子实时（SMRT）基因组和甲基化组测序、亚硫酸氢盐测序，定义了两种最广泛的实验室菌株（中间毕赤酵母ATCC 25611和中间毕赤酵母菌株17）中存在的外源DNA的特异性遗传屏障。我们确定并表征了多个限制修饰（R-M）系统，其中一些在两种菌株之间存在很大差异。我们认为这些R-M系统是中间偃麦草转化障碍的根本原因。此外，我们注意到两种菌株中存在保守的规则间隔短回文重复序列（CRISPR）系统，这可能为外源DNA摄取和掺入提供进一步的屏障。这项工作将为中间疟原虫的遗传系统的开发提供宝贵的资源，这将是对该生物体的生理学，代谢和人类疾病发病机制的基础研究所必需的。
Prevotella intermedia, a major periodontal pathogen, is increasingly implicated in human respiratory tract and cystic fibrosis lung infections. Nevertheless, the specific mechanisms employed by this pathogen remain only partially characterized and poorly understood, largely due to its total lack of genetic accessibility. Here, using Single Molecule, Real-Time (SMRT) genome and methylome sequencing, bisulfite sequencing, in addition to cloning and restriction analysis, we define the specific genetic barriers to exogenous DNA present in two of the most widespread laboratory strains, P. intermedia ATCC 25611 and P. intermedia Strain 17. We identified and characterized multiple restriction-modification (R-M) systems, some of which are considerably divergent between the two strains. We propose that these R-M systems are the root cause of the P. intermedia transformation barrier. Additionally, we note the presence of conserved Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) systems in both strains, which could provide a further barrier to exogenous DNA uptake and incorporation. This work will provide a valuable resource during the development of a genetic system for P. intermedia, which will be required for fundamental investigation of this organism’s physiology, metabolism, and pathogenesis in human disease.
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影响因子：
48
作者：
Flusberg, Benjamin A.;Webster, Dale R.;Lee, Jessica H.;Travers, Kevin J.;Olivares, Eric C.;Clark, Tyson A.;Korlach, Jonas;Turner, Stephen W.
通讯作者：
Turner, Stephen W.
影响因子：
14.9
作者：
Grissa, Ibtissem;Vergnaud, Gilles;Pourcel, Christine
通讯作者：
Pourcel, Christine
DNA 亚硫酸氢盐转化中的错误：调节不适当和失败的转化频率。
影响因子：
14.9
作者：
Genereux DP;Johnson WC;Burden AF;Stöger R;Laird CD
通讯作者：
Laird CD
影响因子：
16.6
作者：
Dupuis, Marie-Eve;Villion, Manuela;Moineau, Sylvain
通讯作者：
Moineau, Sylvain
影响因子：
3.1
作者：
Dorn, BR;Leung, KP;Progulske-Fox, A
通讯作者：
Progulske-Fox, A