Genes of Salmonella enterica Serovar Enteritidis Involved in Biofilm Formation

Seulgi Lee, Jinru Chen
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Abstract

Although biofilms contribute to bacterial tolerance to desiccation and survival in low-moisture foods, the molecular mechanisms underlying biofilm formation have not been fully understood. This study created a mutant library from Salmonella Enteritidis using mini-Tn10 transposon mutagenesis. The biofilm-forming potential of acquired mutants was assessed before the genomic DNA of the mutants that formed significantly (p ≤ 0.05) less biofilm mass than their wildtype parent strain was extracted for deep DNA sequencing. The gene of each mutant interrupted by mini-Tn10 insertion was identified by aligning obtained sequencing data with the reference Genbank sequences using a BLAST search. Sixty-four mutant colonies were selected, and five mutants that formed the least amount of biofilm mass compared to the wildtype parent strain were selected for sequencing analysis. The results of the BLAST search revealed that the gene interrupted by mini-Tn10 in each mutant is responsible for the biosynthesis of aldehyde dehydrogenase (EutE), cysteine desulfurase (SufS or SufE), a transporter protein, porin OmpL, and a ribbon–helix–helix protein from the CopG family, respectively. Knock-off mutant construction is a possible approach to verify the potential of the identified genes to serve as targets of antimicrobial intervention to control Salmonella colonization on low-moisture foods and in their production environment.
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肠炎沙门氏菌参与生物膜形成的基因
虽然生物膜有助于细菌耐受干燥和在低水分食物中存活,但生物膜形成的分子机制尚未完全清楚。本研究利用迷你 Tn10 转座子诱变技术创建了肠炎沙门氏菌突变体库。在对获得的突变体的生物膜形成潜力进行评估之前,先提取生物膜质量明显低于野生型亲本菌株(p ≤ 0.05)的突变体的基因组 DNA 进行深度 DNA 测序。通过使用 BLAST 搜索将获得的测序数据与 Genbank 参考序列进行比对,确定了每个被 Mini-Tn10 插入中断的突变体的基因。筛选出 64 个突变体菌落,并选择与野生型亲本菌株相比形成生物膜量最少的 5 个突变体进行测序分析。BLAST 搜索结果显示,每个突变体中被 mini-Tn10 中断的基因分别负责醛脱氢酶(EutE)、半胱氨酸脱硫酶(SufS 或 SufE)、转运蛋白、孔蛋白 OmpL 和 CopG 家族的带状螺旋-螺旋蛋白的生物合成。构建基因敲除突变体是一种可行的方法,可验证已鉴定基因作为抗菌干预目标的潜力,以控制沙门氏菌在低水分食品及其生产环境中的定植。
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