Role of Biosynthetic Gene Cluster BGC3 in the Cariogenic Virulence of Streptococcus mutans.

Jing Yi Yang, Yi Xin Zhang, Yu Wei Zhang, Ying Chen, Min Di Xu, Dan Dan Wang, Yi Hua Chen, Yi Xiang Wang, Bin Xia
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Abstract

Objective: To investigate the role of the biosynthetic gene cluster BGC3 of Streptococcus mutans (S. mutans) in the process of dental caries.

Methods: BGC3 and ∆BGC3 S. mutans strains were constructed and their growth curves were evaluated. Acid production capacity was assessed by evaluating pH reduction levels over identical culture periods. The survival of bacteria in phosphate citrate buffer solution (pH 3.0) was quantified. The expression levels of virulence genes (atpF, gtfC, gtfD, spaP, vicR and ftf) were analysed using the qPCR. Co-culture experiments were conducted to evaluate bacterial adaptability. Bacterial viability was determined by microscopical examination of live/dead staining.

Results: Deletion of BGC3 did not significantly impact S. mutans growth or acid production in biofilms. The ∆BGC3 strain exhibited enhanced acid resistance and higher expression levels of virulence genes compared to the wild type. In addition, ∆BGC3 exhibited superior bacterial viability in the co-culture system.

Conclusion: BGC3 affected the acid resistance and expression of caries-related genes in S. mutans. The BGC3 knockout strain exhibited a more robust survival capability than the wild-type strain.

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生物合成基因簇BGC3在变形链球菌致龋毒力中的作用。
目的:探讨变形链球菌(S. mutans)生物合成基因簇BGC3在龋发生过程中的作用。方法:构建BGC3和∆BGC3突变链球菌,并对其生长曲线进行评价。通过在相同的培养周期内评估pH降低水平来评估酸生产能力。测定细菌在柠檬酸磷酸盐缓冲液(pH 3.0)中的存活率。采用qPCR分析毒力基因(atpF、gtfC、gtfD、spaP、vicR和ftf)的表达水平。通过共培养实验来评价细菌的适应性。通过活/死染色显微镜检查确定细菌活力。结果:BGC3的缺失对变形链球菌的生长和生物膜的产酸没有显著影响。与野生型相比,∆BGC3菌株表现出更强的耐酸性和更高的毒力基因表达水平。此外,∆BGC3在共培养体系中表现出较好的细菌活力。结论:BGC3影响变形链球菌的耐酸性及龋相关基因的表达。BGC3敲除菌株比野生型菌株表现出更强的生存能力。
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