Transformation and allelic replacement in Francisella spp.

Anthony, L. S. D.; Gu, Minghong; Cowley, Siobhán C.; Leung, Wendy W. S.; Nano, Francis E.

doi:10.1099/00221287-137-12-2697

Cited by 60 publications

(63 citation statements)

References 25 publications

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“…Cas9 regulatory axis deletion mutants and complementation strains were described previously (11,42). FTN_1254 and FTN_0109 mutants were constructed by allelic exchange as described previously (43,44) using primers in SI Appendix, Table S3.…”

Section: Methodsmentioning

confidence: 99%

A CRISPR-Cas system enhances envelope integrity mediating antibiotic resistance and inflammasome evasion

Sampson

Napier

Schroeder³

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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Significance Increasing the integrity of the bacterial envelope is necessary to allow the successful survival of bacterial pathogens within the host and allow them to counteract damage caused by membrane-targeting antibiotics. We demonstrate that components of a clustered, regularly interspaced, short palindromic repeats–CRISPR associated (CRISPR-Cas) system, a prokaryotic defense against viruses and foreign nucleic acid, act to regulate the permeability of the bacterial envelope, ultimately providing these cells with the capability to resist membrane damage caused by antibiotics. This regulation further allows bacteria to resist detection by multiple host receptors to promote virulence. Overall, this study demonstrates the breadth of function of CRISPR-Cas systems in regulation, antibiotic resistance, innate immune evasion, and virulence.

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Section: Methodsmentioning

confidence: 99%

A CRISPR-Cas system enhances envelope integrity mediating antibiotic resistance and inflammasome evasion

Sampson

Napier

Schroeder³

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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“…Chemical transformation of DNA has been used successfully only in F. tularensis subsp. novicida, and efficiency of DNA uptake depends on the plasmid construct and the length and source of DNA Moreover, Francisella DNA is transformed with higher efficiency than non-Francisella DNA (10,123). Attempts to transform LVS using this method were unsuccessful (10).…”

Section: Genetic Tools For Manipulation Of Francisella Tularensismentioning

confidence: 99%

“…novicida, and efficiency of DNA uptake depends on the plasmid construct and the length and source of DNA Moreover, Francisella DNA is transformed with higher efficiency than non-Francisella DNA (10,123). Attempts to transform LVS using this method were unsuccessful (10). Electroporation is an efficient method of introducing DNA into type A Francisella spp., LVS, and F. tularensis subsp.…”

Section: Genetic Tools For Manipulation Of Francisella Tularensismentioning

confidence: 99%

“…Electroporation is an efficient method of introducing DNA into type A Francisella spp., LVS, and F. tularensis subsp. novicida (10,86,100). Transformation efficiency of electroporated DNA is approximately 3-6 × 10 7 CFU g −1 of DNA in LVS and 2 × 10 3 CFU g −1 of DNA in F. tularensis subsp.…”

Section: Genetic Tools For Manipulation Of Francisella Tularensismentioning

confidence: 99%

“…novicida is 2 × 10 8 when novicida DNA is transformed, suggesting the presence of a restriction modification system; no such system has been observed in LVS (86). Subspecies and strains of Francisella transform with different efficiencies and the presence of an exopolysaccharide capsule may affect DNA uptake (10,13,86,100). Cryotransformation is an efficient and reliable method of introducing DNA into F. tularensis subsp.…”

Section: Genetic Tools For Manipulation Of Francisella Tularensismentioning

confidence: 99%

See 2 more Smart Citations

Francisella tularensis:Taxonomy, Genetics, and Immunopathogenesis of a Potential Agent of Biowarfare

McLendon

Apicella

Allen

2006

Annu. Rev. Microbiol.

208

292

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Tularemia is a zoonosis of humans caused by infection with the facultative intracellular bacterium Francisella tularensis. Interest in F. tularensis has increased markedly in the past few years because of its potential use as an agent of bioterrorism. Five subspecies of this organism are found in the Northern hemisphere, but only F. tularensis subsp. tularensis and subsp. holarctica cause disease in humans. This review summarizes what is known about the pathogenesis of tularemia with a focus on bacterial surface components such as lipopolysaccharide and capsule as well as information obtained from the F. tularensis subsp. tularensis SCHU S4 genome. In particular, the mechanisms of action of recently identified virulence factors are discussed in the context of bacterial replication in macrophages and manipulation of the host inflammatory response. Throughout this report shared and unique features of F. tularensis subsp. tularensis, subsp. holarctica, and subsp. novicida are discussed.

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Francisella

Sjöstedt¹

2015

Bergey's Manual of Systematics of Archaea and Bacteria

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Fran.ci.sel' la . M.L. dim. ending ‐ ella ; M.L. fem. n. Francisella named after Edward Francis, American bacteriologist, who extensively studied the etiologic agent and pathogenesis of tularemia and is credited with naming the disease. Proteobacteria / Gammaproteobacteria / Thiotrichales / Francisellaceae / Francisella Short, rod‐shaped or coccoid cells, 0.2–0.7 × 0.2 µm ( Francisella tularensis subsp. tularensis, F. tularensis subsp. holarctica , and F. tularensis subsp. mediasiatica ) or 0.7 × 1.7 µm ( F. philomiragia and F. tularensis subsp. novicida ) when examined during active growth, pleomorphism occurs thereafter. Gram negative, faintly staining. Endospores are not produced. Nonmotile. Aerobic , growth enhanced under microaerophilic conditions. Colonies are distinct, convex, pale white, and reach maximum size in 3–4 days. Cysteine (or cystine) is required for growth ( F. tularensis subsp . tularensis , F. tularensis subsp . holarctica , and F. tularensis subsp . mediasiatica ) or enhances growth ( F. philomiragia and F. tularensis subsp . novicida ). Weakly catalase positive. F. tularensis is oxidase negative and F. philomiragia oxidase positive (Kovacs modification). H 2 S is produced in cysteine‐supplemented medium. Strains from both species, with the exception of F. tularensis subsp. mediasiatica , produce acid but no gas from D ‐glucose and maltose. Urease negative and no reduction of nitrate. Strains are characterized by a unique fatty acid composition ; long‐chain saturated and monosaturated C 18 to C 26 acids, relatively large amounts of saturated even‐chain acids (C 10:0 , C 14:0 , and C 16:0 ), and two long‐chain hydroxy acids (C 16:0 3OH and C 18:0 3OH ). Type species : Francisella tularensis (McCoy and Chapin 1912) Dorofe'ev 1947, 176 ( Bacterium tularense McCoy and Chapin 1912, 61.)

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Transformation and allelic replacement in Francisella spp.

Cited by 60 publications

References 25 publications

A CRISPR-Cas system enhances envelope integrity mediating antibiotic resistance and inflammasome evasion

A CRISPR-Cas system enhances envelope integrity mediating antibiotic resistance and inflammasome evasion

Francisella tularensis:Taxonomy, Genetics, and Immunopathogenesis of a Potential Agent of Biowarfare

Francisella

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