ArticleApplied and environmental microbiology2015
Kanamycin Resistance Cassette for Genetic Manipulation of Treponema denticola.
Article in Applied and environmental microbiology, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed, 14 citations in OpenAlex.
- Expanded Functional Characterization and Optimization of Protein Expression in Treponema denticola Shuttle Plasmids.Molecular oral microbiology · 2026Article
- Enhanced transformation efficiency in Treponema denticola enabled by SyngenicDNA-based plasmids lacking restriction-modification target motifs.Molecular oral microbiology · 2023Article
- Illuminating the oral microbiome and its host interactions: tools and approaches for molecular microbiology studies.FEMS microbiology reviews · 2023Review
- Characterization of Treponema denticola Major Surface Protein (Msp) by Deletion Analysis and Advanced Molecular Modeling.Journal of bacteriology · 2022Article
- Approaches to Understanding Mechanisms of Dentilisin Protease Complex Expression inFrontiers in cellular and infection microbiology · 2021Review
- Genetic Manipulations of Oral Spirochete Treponema denticola.Methods in molecular biology (Clifton, N.J.) · 2021Article
- The Role ofFrontiers in cellular and infection microbiology · 2019Article
- Activation of the Innate Immune System by Treponema denticola Periplasmic Flagella through Toll-Like Receptor 2.Infection and immunity · 2018Article
- 1,2-Diacylglycerol choline phosphotransferase catalyzes the final step in the unique Treponema denticola phosphatidylcholine biosynthesis pathway.Molecular microbiology · 2017Article
- pyrF as a Counterselectable Marker for Unmarked Genetic Manipulations in Treponema denticola.Applied and environmental microbiology · 2016Article
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Authors and funding
3 authors at 1 institution in 1 country.
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Abstract
Treponema denticola has been recognized as an important oral pathogen of the "red complex" bacterial consortium that is associated with the pathogenesis of endodontal and periodontal diseases. However, little is known about the virulence of T. denticola due to its recalcitrant genetic system. The difficulty in genetically manipulating oral spirochetes is partially due to the lack of antibiotic resistance cassettes that are useful for gene complementation following allelic replacement mutagenesis. In this study, a kanamycin resistance cassette was identified and developed for the genetic manipulation of T. denticola ATCC 35405. Compared to the widely used ermF-ermAM cassette, the kanamycin cassette used in the transformation experiments gave rise to additional antibiotic-resistant T. denticola colonies. The kanamycin cassette is effective for allelic replacement mutagenesis as demonstrated by inactivation of two open reading frames of T. denticola, TDE1430 and TDE0911. In addition, the cassette is also functional in trans-chromosomal complementation. This was determined by functional rescue of a periplasmic flagellum (PF)-deficient mutant that had the flgE gene coding for PF hook protein inactivated. The integration of the full-length flgE gene into the genome of the flgE mutant rescued all of the defects associated with the flgE mutant that included the lack of PF filament and spirochetal motility. Taken together, we demonstrate that the kanamycin resistance gene is a suitable cassette for the genetic manipulation of T. denticola that will facilitate the characterization of virulence factors attributed to this important oral pathogen.
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