Molecular analyses and identifications of new genes to infectious diseases relating with oral biofilm

口腔生物膜相关传染病新基因的分子分析及鉴定

基本信息

批准号：
15390571
负责人：
SENPUKU Hidenobu
金额：
$ 9.15万
依托单位：
National Institute of Infectious Diseases
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15390571/
关键词：
mutans streptococci oral biofilm quorum sensing (QS) system DNA micro array Flow cell system bacitracin transport ATP-binding protein bcrA RT-PCR CSP クオラムセンシング(QS)システム Competence stimulating peptide(CSP)ミュータンス連鎖球菌パルスフィールド電気泳動バクテ

项目摘要

A biofilm forming is associated with bacterial pathogenicity and escape from immunological effects in bacterial infectious diseases such as dental caries. Many investigators studied indirectly mechanism of oral biofilm in vitro assay, which is not similar to oral condition. In the present study, we attempted to identify susceptible genes for biofilm formation of Streptococcus mutans in similar condition to oral cavity. Further, we analyzed relationship between the gene expression and host gene (e2f-1) that response to byposalivation in mouse. Comparison of the intensity of biofilm formation by S.mutans clinical strains in flow cell system with the gene expression was firstly investigated. Seventeen different patterns of genomic DNA were identified in S.mutans organisms isolated from biofilm samples taken from 3-year-old children and their mothers by pulsed-field gel electrophoresis. The S.mutans isolates showed differing abilities to form biofilms on flow cell system and the FSC-3 and FSC-4 strains showed the greatest and least, respectively, levels of biofilm formation when examined with confocal laser scanning microscopy (CLSM). Further, in a comparison of FSC-3 and -4 biofilms, DNA microarray results showed 74 genes were differentially expressed more than 4-fold. We identified more than 60% CSP-induced genes of 74 in quorum sensing (QS) system, including bacitracin transport ATP-binding protein bcrA and PTS system IIA, B, C and D. Analyses of the bcrA deficient strains showed significant deceasing of biofilm formation and attachment on the tooth surface in E2F-1^<-/-> mouse in comparison with wild strain. Our results showed that these genes may play biofilm formation and interact with E2F-1 gene in host.

形成的生物膜与细菌的致病性有关，并摆脱细菌感染性疾病（例如龋齿）的免疫作用。许多研究者在体外测定中研究了口服生物膜的间接机制，该机制与口服状况不同。在本研究中，我们试图鉴定出与口腔相似的链球菌突变体生物膜形成的易感基因。此外，我们分析了基因表达与宿主基因（E2F-1）之间对小鼠旁卵裂的反应之间的关系。首先研究了流量细胞系统中S.Mutans临床菌株与基因表达中生物膜形成强度的比较。通过脉冲场凝胶电泳从3岁儿童及其母亲中从3岁儿童及其母亲中分离出的S.Mutans生物中鉴定出了17种不同的基因组DNA模式。 S.Mutans的分离株显示出在流动细胞系统上形成生物膜的能力不同，FSC-3和FSC-4菌株在使用共聚焦激光扫描显微镜（CLSM）检查时，分别显示出最大和最少的生物膜形成水平。此外，在比较FSC-3和-4生物膜时，DNA微阵列结果显示74个基因被差异表达超过4倍以上。我们确定了在法定传感系统中的60％以上CSP诱导的基因，其中74个基因（包括杆状蛋白酶传输ATP结合蛋白BCRA和PTS System IIA，B，C和D与野生应变相比，在E2F-1^< - / - >小鼠中牙齿表面的形成和附着。我们的结果表明，这些基因可能在宿主中发挥生物膜形成并与E2F-1基因相互作用。