Protein synthesis by intact Coxiella burnetii cells

Zuerner, Richard L.; Thompson, Herbert A.

doi:10.1128/jb.156.1.186-191.1983

Cited by 35 publications

(26 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…C. burnetii Nine Mile strain phase I clone 7 (9MIC7) were grown in persistently infected BHK-21 cell lines (ATCC CCL10) as previously described [8]. C. burnetii transformed with ampicillin resistance plasmid pSKO(+)1000 were grown in BHK cells propagated in DMEM containing 10% newborn calf serum with 50 Wg ml 31 ampicillin [6].…”

Section: Bacterial Strains Cells Lines and Growth Conditionsmentioning

confidence: 99%

“…Intracellular and extracellular C. burnetii cells were iso-0378-1097 / 00 / $20.00 ß 2000 Published by Elsevier Science B.V. All rights reserved. PII: S 0 3 7 8 -1 0 9 7 ( 0 0 ) 0 0 0 6 4 -1 lated and partially puri¢ed from host cells as previously described [8]. Organisms were further puri¢ed by centrifugation in celite [9] or in 30% sucrose (w/v), 7.6% renogra¢n in PBS [10].…”

Section: Bacterial Strains Cells Lines and Growth Conditionsmentioning

confidence: 99%

“…Intracellular and extracellular C. burnetii transformants were harvested from host cells, suspended in medium A [8] containing 1 mM MgCl 2 and treated with DNase1 (100 Wg ml 31 ) for 2 h at 37³C. Cells were puri¢ed from host cell debris by di¡erential centrifugation in sucrose/renogra¢n (see above).…”

Section: Sodium Dodecylsulfate-polyacrylamide Gel Electrophoresis (Sdmentioning

confidence: 99%

“…Intracellular and extracellular C. burnetii (9MIC7) cells, grown in BHK-21 cell lines [8], were recovered from cultures and puri¢ed from host cell debris. Competent C. burnetii cells were prepared and electroporated with the ars-bla plasmid pSKO(+)1000 [6] and used to establish persistent infections in BHK-21 ¢broblasts grown in ampicillin (50 Wg ml 31 ).…”

Section: Transformation Of C Burnetiimentioning

confidence: 99%

See 3 more Smart Citations

Expression of β-lactamase in Coxiella burnetii transformants

Suhan¹

2000

FEMS Microbiology Letters

View full text Add to dashboard Cite

Coxiella burnetii can be transformed to ampicillin resistance by electroporation with plasmids encoding L-lactamase. However, nonplasmid emergence of resistance to ampicillin also develops. To validate the usefulness of the bla gene marker for selection and detection, transformed C. burnetii were examined for L-lactamase expression by use of immunoblotting after SDS-PAGE. The 29-kDa mature form of the L-lactamase protein was detected in C. burnetii lysates. Quantitation of these immunoblot signals showed that C. burnetii surprisingly expressed low levels of L-lactamase. The results validate the use of plasmid-encoded ampicillin resistance as a means for selecting C. burnetii transformants; they also suggest that levels of ampicillin used for selection pressure should be empirically determined and that detection of L-lactamase by antibody blotting done to confirm transformants. ß

show abstract

Section: Bacterial Strains Cells Lines and Growth Conditionsmentioning

confidence: 99%

Section: Bacterial Strains Cells Lines and Growth Conditionsmentioning

confidence: 99%

Section: Sodium Dodecylsulfate-polyacrylamide Gel Electrophoresis (Sdmentioning

confidence: 99%

Section: Transformation Of C Burnetiimentioning

confidence: 99%

See 2 more Smart Citations

Expression of β-lactamase in Coxiella burnetii transformants

Suhan¹

2000

FEMS Microbiology Letters

View full text Add to dashboard Cite

show abstract

“…C. burnetii organisms infect a wide variety of host cells in vitro, including human monocytes (THP-1 cells), rodent fibroblasts (L-929 and BHK-21 cells), Vero cells, chick endodermal (primary culture) cells, Chinese hamster ovary cells, and several mouse macrophage cell lines [2,[9][10][11]. In most cell lines that have been used for study, Phase II organisms invade host cells faster than Phase I organisms and are more successful at establishing persistent infections [11].…”

Section: Introductionmentioning

confidence: 99%

A growth study of Coxiella burnetii Nine Mile Phase I and Phase II in fibroblasts

Miller

2004

FEMS Immunology and Medical Microbiology

View full text Add to dashboard Cite

Coxiella burnetii, a slow-growing, gram-negative, obligate intracellular bacterium, is the causative agent of Q fever in humans. The avirulent Phase II C. burnetii Nine Mile strain can invade and establish persistent infections in a wide variety of laboratory cell lines, and is generally considered to be easier to grow in culture than the wild-type Phase I organism. Efforts to improve Phase I organism yield in the BHK-21 cell line demonstrated that high CO2 conditions and the use of Dulbecco's modified Eagle's medium (DMEM) with 4.5 g/l glucose supplementation resulted in higher organism yields. Phase II organisms grown in the same cell line and conditions showed lower growth rates. Analysis revealed that increased average numbers of C. burnetii Phase I organisms within fibroblasts was due to higher growth rates within the hosts rather than to increased uptake or to increased cell-to-cell spreading. Addition of the nucleoside cytidine to the growth medium stimulated growth of Phase II but not Phase I organisms.

show abstract

Coxiella

Drancourt¹,

Raoult²

2015

Bergey's Manual of Systematics of Archaea and Bacteria

View full text Add to dashboard Cite

Co.xi.el' la . M.L. fem. dim. ‐ ella ending; M.L. fem. n. Coxiella named after Harold R. Cox, who in collaboration with G.E. Davis, first isolated this organism in the United States shortly after its discovery in Australia and who introduced the technique of yolk sac inoculation of the chick embryo, which greatly facilitated the study of this and other genera. Proteobacteria / Gammaproteobacteria / Legionellales / Coxiellaceae / Coxiella Strictly intracellular bacteria , usually 0.2–0.4 µm × 0.4–1.0 µm. Best stained by Gimenez staining . They have no flagella or capsule. They live in close natural association with arthropod and vertebrate hosts . The genus includes Coxiella burnetii —the agent of Q fever —and endosymbionts of ticks and aquatic invertebrates. C. burnetii grows well in cultured cell lines and in the yolk sac of chicken embryos, where it undergoes a cycle of development with formation of an endospore‐like body. Coxiella burnetii possesses two antigenic phases : natural virulent phase I and attenuated phase II, which is obtained after passage in cultured cell lines and yolk sac. The organism grows in vacuoles of the host cell rather than in the cytoplasm or the nucleus. C. burnetii phase I vacuoles share characteristics of late‐stage, immature phagosomes whereas C. burnetii phase II vacuoles are bactericidal phagolysosomes. Highly resistant to chemical agents and elevated temperature. Coxiellae occur worldwide in ticks and various vertebrates, including humans. Infection is particularly prevalent in cattle, sheep and goats. Extremely infectious; a single aerosol‐borne organism can cause the human disease Q fever. C. burnetii is one of the ten most feared potential bioterrorism agents . The mol % G + C of the DNA is : 42.7. Type species : Coxiella burnetii (Derrick 1939) Philip 1948, 58 AL ( Rickettsia burnetii (sic) Derrick 1939, 14.)

show abstract

Protein synthesis by intact Coxiella burnetii cells

Cited by 35 publications

References 18 publications

Expression of β-lactamase in Coxiella burnetii transformants

Expression of β-lactamase in Coxiella burnetii transformants

A growth study of Coxiella burnetii Nine Mile Phase I and Phase II in fibroblasts

Coxiella

Contact Info

Product

Resources

About