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在不同蔗糖浓度条件下对变形链球菌中蔗糖诱导的小RNA的分析。

Analysis of sucrose-induced small RNAs in Streptococcus mutans in the presence of different sucrose concentrations.

作者信息

Liu Shan Shan, Zhu Wen Hui, Zhi Qing Hui, Liu Jia, Wang Yan, Lin Huan Cai

机构信息

Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-Sen University, Guangdong Provincial Key Laboratory of Stomatology, Guangzhou, 510055, China.

出版信息

Appl Microbiol Biotechnol. 2017 Jul;101(14):5739-5748. doi: 10.1007/s00253-017-8346-x. Epub 2017 May 31.

Abstract

Streptococcus mutans (S. mutans) is the major pathogen contributing to dental caries. Sucrose is an important carbohydrate source for S. mutans and is crucial for dental caries. Small RNAs (sRNAs) are key post-transcriptional regulators of stress adaptation and virulence in bacteria. Here, for the first time, we created three replicate RNA libraries exposed to either 1 or 5% sucrose. The expression levels of sRNAs and target genes (gtfB, gtfC, and spaP) related to virulence were assessed. In addition, some phenotypic traits were evaluated. We obtained 2125 sRNA candidates with at least 100 average reads in 1% sucrose or 5% sucrose. Of these candidates, 2 were upregulated and 20 were downregulated in 1% sucrose. Six of these 22 differentially expressed sRNAs were validated by qRT-PCR. The expression level of target gene gtfB was higher in 1% sucrose. The adherence ratio of S. mutans was higher in 1% sucrose than in 5% sucrose. The synthesis of water-insoluble glucans (WIGs) was significantly higher in 5% sucrose than in 1% sucrose. These data suggest that a series of sRNAs can be induced in response to sucrose, and that some sRNAs might be involved in the regulation of phenotypes, providing new insight into the prevention of caries.

摘要

变形链球菌是导致龋齿的主要病原体。蔗糖是变形链球菌重要的碳水化合物来源,对龋齿形成至关重要。小RNA(sRNA)是细菌应激适应和毒力的关键转录后调节因子。在此,我们首次创建了三个分别暴露于1%或5%蔗糖的重复RNA文库。评估了与毒力相关的sRNA和靶基因(gtfB、gtfC和spaP)的表达水平。此外,还评估了一些表型特征。我们在1%或5%蔗糖中获得了2125个平均读数至少为100的sRNA候选物。在这些候选物中,2个在1%蔗糖中上调,20个在1%蔗糖中下调。通过qRT-PCR验证了这22个差异表达的sRNA中的6个。靶基因gtfB在1%蔗糖中的表达水平更高。变形链球菌在1%蔗糖中的黏附率高于在5%蔗糖中的黏附率。5%蔗糖中不溶性葡聚糖(WIGs)的合成明显高于1%蔗糖。这些数据表明,一系列sRNA可响应蔗糖诱导产生,且一些sRNA可能参与表型调控,为龋齿预防提供了新见解。

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