Salmonella is a bacterial pathogen frequently involved in human gastrointestinal infections including those associated with low-moisture foods such as dehydrated food powders/spices, vegetable seeds, and tree nuts. The survival/persistence of Salmonella on low moisture foods and in dry environments is enhanced by its ability in developing biofilms. This study was undertaken to identify the genetic determinants critical for Salmonella attachment and biofilm formation. E. coli SM10 lambda pir, with a kanamycin resistant marker on mini-Tn10 (mini-Tn10:lacZ:kanr), an ampicillin resistant marker on the mini-Tn10-bearing suicidal plasmid pLBT and a streptomycin sensitive marker on the SM10 chromosome, was used as a donor (ampr, kanr, streps), and three Salmonella strains (amps, kans, strepr) were used as recipients in a transposon mutagenesis study. The donor and each recipient were co-incubated overnight on tryptic soy agar at 37 °C, and mutant colonies (amps, kanr, strepr) were subsequently selected. A single-banded degenerate PCR product, amplified from each mutant genome using oligonucleotide primers derived from the end of min-Tn10 and restriction enzyme EcoR I- or Pst I-recognizing sequence, were analyzed using the Sanger sequencing technology. Acquired DNA sequences were compared to those deposited in the Genbank using BLAST search. Cells of Salmonella mutants accumulated either significantly more or less (P < 0.05) biofilms than their parent cells on polystyrene surface. Sequence analysis of degenerate PCR products revealed that the mini-Tn10 from pLBT had inserted into the cdg, trx, fadI or rxt on Salmonella chromosomes. Results of the research will likely help strategize future antimicrobial intervention for control of pathogen attachment and biofilm formation.

中文翻译：

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肠沙门氏菌的遗传决定因素对于附着和生物膜形成至关重要。

沙门氏菌是一种细菌病原体，经常参与人类胃肠道感染，包括与低水分食物（如脱水食物粉/香料，蔬菜种子和坚果）相关的感染。沙门氏菌在生物膜上的形成能力增强了沙门氏菌在低水分食品和干燥环境中的生存/持久性。进行这项研究以鉴定对沙门氏菌附着和生物膜形成至关重要的遗传决定因素。大肠杆菌SM10 lambda pir，在mini-Tn10上具有卡那霉素抗性标记（mini-Tn10：lacZ：kanr），在带有mini-Tn10的自杀质粒pLBT上具有氨苄青霉素抗性标记，在SM10染色体上具有链霉素敏感标记，转座子诱变研究使用三株沙门氏菌（amps，kans，strepr）作为供体（ampr，kanr，streps），并使用三种沙门氏菌菌株（amps，kans，strepr）作为受体。供体和每个受体在胰蛋白酶大豆琼脂上于37°C共孵育过夜，随后选择突变菌落（amps，kanr，strepr）。使用Sanger测序技术分析了使用min-Tn10末端的寡核苷酸引物和限制性酶EcoR I-或Pst I识别序列从每个突变基因组扩增的单带简并PCR产物。使用BLAST搜索将获得的DNA序列与Genbank中保存的DNA序列进行比较。沙门氏菌突变体的细胞比其亲代细胞在聚苯乙烯表面上积聚了更多或更少（P <0.05）的生物膜。简并PCR产物的序列分析表明，来自pLBT的mini-Tn10已插入沙门氏菌染色体上的cdg，trx，fadI或rxt。