Open Access Highly Accessed Research article

Sec-mediated secretion by Coxiella burnetii

Christopher M Stead1, Anders Omsland2, Paul A Beare1, Kelsi M Sandoz1 and Robert A Heinzen1*

Author Affiliations

1 Coxiella Pathogenesis Section, Laboratory of Intracellular Parasites, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT 59840, USA

2 Host-Parasite Interactions Section, Laboratory of Intracellular Parasites, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT 59840, USA

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BMC Microbiology 2013, 13:222  doi:10.1186/1471-2180-13-222

Published: 5 October 2013

Additional files

Additional file 1:

Peptide fragments identified in C. burnetii ACCM culture supernatants by microcapillary HPLC, nano-ESI, MS/MS analysis.

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Additional file 2:

List of C. burnetii potentially secreted proteins.

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Additional file 3:

Expression of FLAG-tagged secretion candidates by C. burnetii transformants to confirm secretion.C. burnetii transformed with plasmids encoding FLAG-tagged secretion candidates were cultured for 48 h, then expression of tagged protein induced by addition of aTc for 24 h. Supernatants were harvested, TCA precipitated and analyzed by immunoblotting using antibody directed against the FLAG-tag. Supernatants of samples that were positive for secretion were then probed using antibody directed against EF-Ts to rule out cell lysis as a source of protein present in supernatants. Whole cell lysate of C. burnetii expressing FLAG-tagged CBU1764a was used as a positive control (+ve). To confirm that proteins not present in supernatants were expressed by C. burnetii transformants, lysates of bacterial pellets were probed with antibody directed against the FLAG-tag.

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Additional file 4:

Comparison of F. novicida and C. burnetii pil genes. The C. burnetii genome contains 13 pil genes, 11 of which are also present in the F. novicida genome, a bacterium that employs T4P-mediated secretion.

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Additional file 5:

C. burnetii is not pilliated. Transmission electron micrographs of negatively stained bacteria show pili on F. tularensis LVS (panel A) but not C. burnetii (panel B). Scale bars = 0.5 μm.

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Additional file 6:

Primers used in this study.

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