Construction of a New Phage Integration Vector pFIV-Val for Use in Different Francisella Species

Tlapák, Hana; Köppen, Kristin; Rydzewski, Kerstin; Grunow, Roland; Heuner, Klaus

doi:10.3389/fcimb.2018.00075

Cited by 10 publications

(7 citation statements)

References 39 publications

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“…This would be a fruitful undertaking, as it could be shown that F-W12 is genetically treatable (generation of mutant strains by a transposon or of specific mutants by site-specific recombination after natural transformation; transformation of plasmids by electroporation). Recently, a new phage integration vector (pFIV-Val) was built, which can be used for integrating genes into the genome of F-W12, as well as for the complementation of specific mutant strains [ 80 , 81 , 82 ].…”

Section: Francisella Sp Strain W12-1067 (F-w12mentioning

confidence: 99%

Francisella tularensis Subspecies holarctica and Tularemia in Germany

et al. 2020

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Tularemia is a zoonotic disease caused by Francisella tularensis a small, pleomorphic, facultative intracellular bacterium. In Europe, infections in animals and humans are caused mainly by Francisella tularensis subspecies holarctica. Humans can be exposed to the pathogen directly and indirectly through contact with sick animals, carcasses, mosquitoes and ticks, environmental sources such as contaminated water or soil, and food. So far, F. tularensis subsp. holarctica is the only Francisella species known to cause tularemia in Germany. On the basis of surveillance data, outbreak investigations, and literature, we review herein the epidemiological situation—noteworthy clinical cases next to genetic diversity of F. tularensis subsp. holarctica strains isolated from patients. In the last 15 years, the yearly number of notified cases of tularemia has increased steadily in Germany, suggesting that the disease is re-emerging. By sequencing F. tularensis subsp. holarctica genomes, knowledge has been added to recent findings, completing the picture of genotypic diversity and geographical segregation of Francisella clades in Germany. Here, we also shortly summarize the current knowledge about a new Francisella species (Francisella sp. strain W12-1067) that has been recently identified in Germany. This species is the second Francisella species discovered in Germany.

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Section: Francisella Sp Strain W12-1067 (F-w12mentioning

confidence: 99%

Francisella tularensis Subspecies holarctica and Tularemia in Germany

et al. 2020

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“…Two hypothetical proteins (Fhv_0032/49) and two integrases (Fhv_0033/51) exhibit homologs (29–62% identity) to proteins in Fno , Fph and F. salina . The protein Fhv_0051 has been identified earlier to be the site-specific integrase of FhaGI-1, necessary for the integration and excision of the phage integration vector pFIV-Val and thus probably also for the prophage [ 19 ].…”

Section: Resultsmentioning

confidence: 99%

“…Francisella strains were cultivated at 37 °C in medium T (MT) (1% brain heart infusion broth (Difco Laboratories, Inc., Sparks, MD, USA), 1% bacto tryptone (Difco Laboratories, Inc., Sparks, MD, USA), 1% technical casamino acids (Difco Laboratories, Inc., Sparks, MD, USA), 0.005% of MgSO 4 , 0.01% FeSO 4 , 0.12% sodium citrate, 0.02% KCl, 0.04% K 2 HPO 4 , 0.06% L-cysteine and 1.5% glucose) [ 32 , 33 ] or on MT agar plates supplemented with hemoglobin and charcoal (MTKH plates, [ 19 ]). E. coli strains were grown at 37 °C in lysogeny broth (LB; 1% bacto tryptone, 0.5% yeast extract, 0.5% NaCl) or on LB agar (LB supplemented with 1.2% agar).…”

Section: Methodsmentioning

confidence: 99%

“…Generation of this episomal form depend on the presence of a site-specific integrase/recombinase [ 11 , 18 ]. The repeat region ( attP ) and the integrase ( int ) of FhaGI-1 were then used to generate a new Francisella integration vector (pFIV-Val) which integrates site-specifically into the tRNA-Val gene of different Francisella species [ 19 ].…”

Section: Introductionmentioning

confidence: 99%

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First Description of a Temperate Bacteriophage (vB_FhiM_KIRK) of Francisella hispaniensis Strain 3523

Köppen

Prensa

Rydzewski

et al. 2021

Viruses

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Here we present the characterization of a Francisella bacteriophage (vB_FhiM_KIRK) including the morphology, the genome sequence and the induction of the prophage. The prophage sequence (FhaGI-1) has previously been identified in F. hispaniensis strain 3523. UV radiation induced the prophage to assemble phage particles consisting of an icosahedral head (~52 nm in diameter), a tail of up to 97 nm in length and a mean width of 9 nm. The double stranded genome of vB_FhiM_KIRK contains 51 open reading frames and is 34,259 bp in length. The genotypic and phylogenetic analysis indicated that this phage seems to belong to the Myoviridae family of bacteriophages. Under the conditions tested here, host cell (Francisella hispaniensis 3523) lysis activity of KIRK was very low, and the phage particles seem to be defective for infecting new bacterial cells. Nevertheless, recombinant KIRK DNA was able to integrate site-specifically into the genome of different Francisella species after DNA transformation.

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“…Transduction: Transduction is the transfer of DNA to a bacterium via a bacteriophage. There have been only a few reports of bacteriophage which can act against Francisella so far ( Koliaditskaia et al, 1959 ; Tlapak et al, 2018 ) and they appear so far to be unstable or difficult to isolate. This area of research is ongoing, but one very interesting finding was the identification of CRISPR/CAS9 system in F. novicida ( Schunder et al, 2013 ), suggesting that there have been bacteriophage interactions with Francisella in the past, enough to acquire a viral immunity system such as CRISPR/Cas.…”

Section: Horizontal Gene Transfer In Francisellamentioning

confidence: 99%

Genetic Determinants of Antibiotic Resistance in Francisella

Kassinger

Hoek

2021

Front. Microbiol.

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Tularemia, caused by Francisella tularensis, is endemic to the northern hemisphere. This zoonotic organism has historically been developed into a biological weapon. For this Tier 1, Category A select agent, it is important to expand our understanding of its mechanisms of antibiotic resistance (AMR). Francisella is unlike many Gram-negative organisms in that it does not have significant plasmid mobility, and does not express AMR mechanisms on plasmids; thus plasmid-mediated resistance does not occur naturally. It is possible to artificially introduce plasmids with AMR markers for cloning and gene expression purposes. In this review, we survey both the experimental research on AMR in Francisella and bioinformatic databases which contain genomic and proteomic data. We explore both the genetic determinants of intrinsic AMR and naturally acquired or engineered antimicrobial resistance as well as phenotypic resistance in Francisella. Herein we survey resistance to beta-lactams, monobactams, carbapenems, aminoglycosides, tetracycline, polymyxins, macrolides, rifampin, fosmidomycin, and fluoroquinolones. We also highlight research about the phenotypic AMR difference between planktonic and biofilm Francisella. We discuss newly developed methods of testing antibiotics against Francisella which involve the intracellular nature of Francisella infection and may better reflect the eventual clinical outcomes for new antibiotic compounds. Understanding the genetically encoded determinants of AMR in Francisella is key to optimizing the treatment of patients and potentially developing new antimicrobials for this dangerous intracellular pathogen.

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Construction of a New Phage Integration Vector pFIV-Val for Use in Different Francisella Species

Cited by 10 publications

References 39 publications

Francisella tularensis Subspecies holarctica and Tularemia in Germany

Francisella tularensis Subspecies holarctica and Tularemia in Germany

First Description of a Temperate Bacteriophage (vB_FhiM_KIRK) of Francisella hispaniensis Strain 3523

Genetic Determinants of Antibiotic Resistance in Francisella

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