ArticleJournal of the Formosan Medical Association = Taiwan yi zhi2014
A markerless protocol for genetic analysis of Aggregatibacter actinomycetemcomitans.
Article in Journal of the Formosan Medical Association = Taiwan yi zhi, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
6 citing papers in PubMed.
- Article
- Genomic integration and expression of the Aggregatibacter actinomycetemcomitans catalase gene in Aggregatibacter aphrophilus.Archives of oral biology · 2018Article
- Article
- The in vivo T helper type 17 and regulatory T cell immune responses to Aggregatibacter actinomycetemcomitans.Molecular oral microbiology · 2017Article
- Article
- Versatile Vectors for Efficient Mutagenesis of Bradyrhizobium diazoefficiens and Other Alphaproteobacteria.Applied and environmental microbiology · 2016Article
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Authors and funding
6 authors.
Funding
Abstract
background/purposeThe genomes of different Aggregatibacter actinomycetemcomitans (A actinomycetemcomitans) strains contain many strain-specific genes and genomic islands (defined as DNA found in some but not all strains) of unknown functions. Genetic analysis for the functions of these islands will be constrained by the limited availability of genetic markers and vectors for A actinomycetemcomitans. In this study, we tested a novel genetic approach of gene deletion and restoration in a naturally competent A actinomycetemcomitans strain D7S-1.
methodsSpecific genes' deletion mutants and mutants restored with the deleted genes were constructed by a markerless loxP/Cre system. In mutants with sequential deletion of multiple genes loxP with different spacer regions were used to avoid unwanted recombinations between loxP sites.
resultsEight single-gene deletion mutants, four multiple-gene deletion mutants, and two mutants with restored genes were constructed. No unintended non-specific deletion mutants were generated by this protocol. The protocol did not negatively affect the growth and biofilm formation of A actinomycetemcomitans.
conclusionThe protocol described in this study is efficient and specific for genetic manipulation of A actinomycetemcomitans, and will be amenable for functional analysis of multiple genes in A actinomycetemcomitans.
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