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利用DNA微阵列技术鉴定与变形链球菌生物膜厚度相关的基因。

DNA-microarrays identification of Streptococcus mutans genes associated with biofilm thickness.

作者信息

Shemesh Moshe, Tam Avshalom, Kott-Gutkowski Miriam, Feldman Mark, Steinberg Doron

机构信息

Biofilm Research Laboratory, Institute of Dental Sciences, Faculty of Dental Medicine, Hebrew University-Hadassah, POB 12272, Jerusalem 91120, Israel.

出版信息

BMC Microbiol. 2008 Dec 29;8:236. doi: 10.1186/1471-2180-8-236.

Abstract

BACKGROUND

A biofilm is a complex community of microorganisms that develop on surfaces in diverse environments. The thickness of the biofilm plays a crucial role in the physiology of the immobilized bacteria. The most cariogenic bacteria, mutans streptococci, are common inhabitants of a dental biofilm community. In this study, DNA-microarray analysis was used to identify differentially expressed genes associated with the thickness of S. mutans biofilms.

RESULTS

Comparative transcriptome analyses indicated that expression of 29 genes was differentially altered in 400- vs. 100-microns depth and 39 genes in 200- vs. 100-microns biofilms. Only 10 S. mutans genes showed differential expression in both 400- vs. 100-microns and 200- vs. 100-microns biofilms. All of these genes were upregulated. As sucrose is a predominant factor in oral biofilm development, its influence was evaluated on selected genes expression in the various depths of biofilms. The presence of sucrose did not noticeably change the regulation of these genes in 400- vs. 100-microns and/or 200- vs. 100-microns biofilms tested by real-time RT-PCR. Furthermore, we analyzed the expression profile of selected biofilm thickness associated genes in the luxS- mutant strain. The expression of those genes was not radically changed in the mutant strain compared to wild-type bacteria in planktonic condition. Only slight downregulation was recorded in SMU.2146c, SMU.574, SMU.609, and SMU.987 genes expression in luxS- bacteria in biofilm vs. planktonic environments.

CONCLUSION

These findings reveal genes associated with the thickness of biofilms of S. mutans. Expression of these genes is apparently not regulated directly by luxS and is not necessarily influenced by the presence of sucrose in the growth media.

摘要

背景

生物膜是在各种环境中的表面上形成的微生物复杂群落。生物膜的厚度在固定化细菌的生理学中起着关键作用。最具致龋性的细菌——变形链球菌,是牙生物膜群落的常见成员。在本研究中,DNA微阵列分析用于鉴定与变形链球菌生物膜厚度相关的差异表达基因。

结果

比较转录组分析表明,在深度为400微米与100微米的生物膜中,29个基因的表达有差异改变;在深度为200微米与100微米的生物膜中,39个基因有差异改变。在深度为400微米与100微米以及200微米与100微米的生物膜中,只有10个变形链球菌基因显示出差异表达。所有这些基因均上调。由于蔗糖是口腔生物膜形成中的主要因素,因此评估了其对生物膜不同深度中选定基因表达的影响。通过实时逆转录聚合酶链反应检测,蔗糖的存在并未显著改变在深度为400微米与100微米和/或200微米与100微米的生物膜中这些基因的调控。此外,我们分析了luxS突变株中选定的与生物膜厚度相关基因的表达谱。与浮游状态下的野生型细菌相比,这些基因在突变株中的表达没有发生根本性变化。在生物膜与浮游环境中,luxS缺陷型细菌的SMU.2146c、SMU.574、SMU.609和SMU.987基因表达仅略有下调。

结论

这些发现揭示了与变形链球菌生物膜厚度相关的基因。这些基因的表达显然不受luxS直接调控,也不一定受生长培养基中蔗糖存在的影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4059/2647549/87e606279365/1471-2180-8-236-1.jpg

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