A mariner transposon vector adapted for mutagenesis in oral streptococci

Nilsson, Marie; Christiansen, Natalia; Høiby, Niels; Twetman, Svante; Givskov, Michael; Tolker‐Nielsen, Tim

doi:10.1002/mbo3.171

Cited by 14 publications

(20 citation statements)

References 36 publications

(61 reference statements)

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“…Though the mariner family of transposons had been used in Gram-positive bacterial mutants such as Streptococcus mutans and Streptococcus pyogenes 36373839 for many years, but this system has not previously been used to study swine streptococcosis pathogens. Himar1 has a ubiquitous dinucleotide target, TA, and requires no species-specific host factors40.…”

Section: Discussionmentioning

confidence: 99%

A novel suicide shuttle plasmid for Streptococcus suis serotype 2 and Streptococcus equi ssp. zooepidemicus gene mutation

Zhang

et al. 2016

Sci Rep

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The mariner-based Himar1 system has been utilized for creating mutant libraries of many Gram-positive bacteria. Streptococcus suis serotype 2 (SS2) and Streptococcus equi ssp. zooepidemicus (SEZ) are primary pathogens of swine that threaten the swine industry in China. To provide a forward-genetics technology for finding virulent phenotype-related genes in these two pathogens, we constructed a novel temperature-sensitive suicide shuttle plasmid, pMar4s, which contains the Himar1 system transposon, TnYLB-1, and the Himar1 C9 transposase from pMarA and the repTAs temperature-sensitive fragment from pSET4s. The kanamycin (Kan) resistance gene was in the TnYLB-1 transposon. Temperature sensitivity and Kan resistance allowed the selection of mutant strains and construction of the mutant library. The SS2 and SEZ mutant libraries were successfully constructed using the pMar4s plasmid. Inverse-Polymerase Chain Reaction (Inverse-PCR) results revealed large variability in transposon insertion sites and that the library could be used for phenotype alteration screening. The thiamine biosynthesis gene apbE was screened for its influence on SS2 anti-phagocytosis; likewise, the sagF gene was identified to be a hemolytic activity-related gene in SEZ. pMar4s was suitable for mutant library construction, providing more information regarding SS2 and SEZ virulence factors and illustrating the pathogenesis of swine streptococcosis.

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Section: Discussionmentioning

confidence: 99%

A novel suicide shuttle plasmid for Streptococcus suis serotype 2 and Streptococcus equi ssp. zooepidemicus gene mutation

Zhang

et al. 2016

Sci Rep

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“…Double function of enolase may explain the results demonstrated almost a twofold increase in the expression of the genes for enolase during the initiation of bacterial biofilm formation. Due to the presence of surface enzymes on the bacterial cells, S. mutans is considered, next to the streptococcus of viridans group, as one of the etiological factors of bacterial endocarditis or bacteremia [82,83]. Also rare cases of recurrent bacteremia in women with Sjogren's syndrome were noted [84].…”

Section: Carbohydrate Metabolismmentioning

confidence: 99%

“…Bacteria present in mixed biofilm can not only interfere with each other on the changes in gene expression, but they may also provide each other with plasmids, e.g., antibiotic resistance genes [78]. Among the S. mutans species, inter alia, erythromycin or kanamycin resistance gene (gene aphA3) can be transferred, which causes the changes in above bacterial phenotype in the direction of multidrug resistance phenotype, impeding an effective targeted therapy [82,107]. An example of such plasmids may be a new transposon vector called "pMN100," containing, among others, selective kanamycin resistance gene, an aminoglycoside antibiotic [82].…”

Section: Mixed Biofilmmentioning

confidence: 99%

“…Among the S. mutans species, inter alia, erythromycin or kanamycin resistance gene (gene aphA3) can be transferred, which causes the changes in above bacterial phenotype in the direction of multidrug resistance phenotype, impeding an effective targeted therapy [82,107]. An example of such plasmids may be a new transposon vector called "pMN100," containing, among others, selective kanamycin resistance gene, an aminoglycoside antibiotic [82]. As the multi-species biofilm develops, S. mutans increases the expression of genes related to the synthesis, alterations, and adhesion of EPS, particularly genes for surface enzymes, glycosyltransferases.…”

Section: Mixed Biofilmmentioning

confidence: 99%

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The Role of Human Oral Microbiome in Dental Biofilm Formation

Krzyściak¹,

Jurczak²,

Piątkowski³

2016

Microbial Biofilms - Importance and Applications

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Each surface of the human body, which stays in contact with the external environment, is covered by a layer of microorganisms. This layer-the human microbiomeis characterized by a high diversity of species and huge number of cells. Its name was proposed by Joshua Lederberg at the turn of the twentieth and twenty-first centuries and was originally referred to as a group of microorganisms colonizing a certain habitat. Currently, the term also defines a set of genomes of all organisms inhabiting a particular niche. Since the human microbiota affects many aspects of human health, it has become the subject of different studies. The use of sequencing methods enabled to obtain genetic material derived directly from the human environment with simultaneous explanation of mutual relationships between microorganisms inhabiting different ecological niches of human organism (i.e., commensal, symbiotic, and pathogenic microorganisms). It is hard to determine the amount of microbiota inhabiting human oral cavity because microbiota represents distinct anatomically limited ecological niches; for example, microbiota of tongue surface, cheek, teeth, palate, gingiva, and periodontal pocket. Apart from anatomical structure, other factors determine different composition of particular oral cavity microbiota. These factors are various qualities of saliva-a natural protective barrier ensuring maintenance of healthy condition of the oral cavity-and habits of diet and hygiene. Generally, bacteria are passively transported by flowing saliva toward teeth surfaces. In turn, the pioneering microorganisms initiating changes in the environment of oral cavity through the production and secretion of products of their metabolism induce mutual microbiota-biofilm interactions. The formation of biofilm of the plaque is a complex and rapidly evolving process in which several stages of development can be distinguished arbitrarily: (i) reversible binding of bacteria to solid surfaces, (ii) production of exopolysaccharide matrix, (iii) irreversible binding to the surface, (iv) maturation of biofilm structure, (v) disintegration and dispersion of an organized structure, and (vi) the formation of new habitats. An oral microbiome

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“…The mariner family of transposons, first identified in horn fly, is known to be widespread in nature. The Himar1 mariner transposon is one of the two known active mariner elements that have been successfully utilized for a number of oral and non-oral bacteria such as Pseudomonas aeruginosa ( Withers et al, 2014 ), Porphyromonas gingivalis ( Klein et al, 2012 ), and oral streptococci ( Nilsson et al, 2014 ). A key feature that separates these elements from many other transposons is their independence from host-specific factors other than the presence of a TA dinucleotide target ( Lampe et al, 1998 ).…”

Section: Introductionmentioning

confidence: 99%

Himar1 Transposon for Efficient Random Mutagenesis in Aggregatibacter actinomycetemcomitans

Ding

Tan

2017

Front. Microbiol.

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Aggregatibacter actinomycetemcomitans is the primary etiological agent of aggressive periodontal disease. Identification of novel virulence factors at the genome-wide level is hindered by lack of efficient genetic tools to perform mutagenesis in this organism. The Himar1 mariner transposon is known to yield a random distribution of insertions in an organism’s genome with requirement for only a TA dinucleotide target and is independent of host-specific factors. However, the utility of this system in A. actinomycetemcomitans is unknown. In this study, we found that Himar1 transposon mutagenesis occurs at a high frequency (×10-4), and can be universally applied to wild-type A. actinomycetemcomitans strains of serotypes a, b, and c. The Himar1 transposon inserts were stably inherited in A. actinomycetemcomitans transconjugants in the absence of antibiotics. A library of 16,000 mutant colonies of A. actinomycetemcomitans was screened for reduced biofilm formation. Mutants with transposon inserts in genes encoding pilus, putative ion transporters, multidrug resistant proteins, transcription regulators and enzymes involved in the synthesis of extracellular polymeric substance, bacterial metabolism and stress response were discovered in this screen. Our results demonstrated the utility of the Himar1 mutagenesis system as a novel genetic tool for functional genomic analysis in A. actinomycetemcomitans.

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A mariner transposon vector adapted for mutagenesis in oral streptococci

Cited by 14 publications

References 36 publications

A novel suicide shuttle plasmid for Streptococcus suis serotype 2 and Streptococcus equi ssp. zooepidemicus gene mutation

A novel suicide shuttle plasmid for Streptococcus suis serotype 2 and Streptococcus equi ssp. zooepidemicus gene mutation

The Role of Human Oral Microbiome in Dental Biofilm Formation

Himar1 Transposon for Efficient Random Mutagenesis in Aggregatibacter actinomycetemcomitans

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