Identification of the Genes Involved in the Biofilm-like Structures on Actinomyces oris K20, a Clinical Isolate from an Apical Lesion
Identification of the Genes Involved in the Biofilm-like Structures on Actinomyces oris K20, a Clinical Isolate from an Apical Lesion
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DOI：
10.1016/j.joen.2012.08.009
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发表时间：
2013-01-01
影响因子：
4.2
通讯作者：
Fukushima, Hisanori
中科院分区：
文献类型：
--
作者：
Mashimo, Chiho;Kamitani, Hiroyuki;Fukushima, Hisanori
简介：虽然生物膜的产生被认为是口腔驻留微生物引起的脓肿形成的发病机制中的关键，但具体机制仍然未知。本研究的目的是鉴定口腔放线菌K20临床分离株中负责维持细胞表面相关网状结构的基因，这些结构存在于一些生物膜产生细菌中。方法：利用转座子EZ-Tn5对K20菌株进行随机插入突变。转座子插入突变体进行了筛选，通过扫描电子显微镜细胞表面相关的网状结构的情况下。通过使用转座子末端引物的直接基因组测序来确定通过转座子插入破坏的基因。结果：从175个突变体中筛选出5个缺失网状结构的突变体。对转座子插入侧翼区的测序结果表明，3个突变体分别具有编码多糖、脱乙酰酶、Spo0J的ParB样核酸酶结构域和假想蛋白的基因。另外2个突变体分别在非编码区和未知区插入。结论：这些基因可能与口腔放线菌K20的网状结构形成有关。（J Endod 2013; 39：44 - 48）。
Introduction: Although the production of biofilm is thought to be crucial in the pathogenesis of abscess formations caused by oral resident microorganisms, the particular mechanisms are still unknown. The aim of this study was to identify gene(s) responsible for maintaining the cell surface-associated meshwork-like structures, which are found in some biofilm-producing bacteria, in a clinical isolate of Actinomyces oris K20. Methods: Random insertional mutagenesis by using transposon EZ-Tn5 was, performed against the strain K20. Transposon insertion mutants were screened by scanning electron microscopy for the absence of cell surface-associated meshwork-like structures. The disrupted genes by the transposon insertion were determined by direct genome sequencing with the transposon-end primers. Results: Five mutants without the meshwork-like structures were identified from 175 mutants. Sequencing of flanking regions of transposon insertion revealed that 3 mutants had a gene encoded polysaccharide, deacetylase, Spo0J containing ParB-like nuclease domain, and hypothetical protein, respectively. The other 2 mutants had an insertion in a noncoding region and an unidentified region respectively. Conclusions: Our findings indicated that these genes might be involved in the formation of meshwork-like structures on Actinomyces oris, K20. (J Endod 2013;39:44-48).