Expression of FLAG-tagged secretion candidates by C. burnetii transformants confirms secretion and not cell lysis.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. Immunoblots were also probed with antibody directed against the cytosolic protein EF-Ts to control for bacterial lysis. Whole cell lysate of C. burnetii expressing FLAG-tagged CBU1764a was used as a positive control (+ve).
Stead et al. BMC Microbiology 2013 13:222 doi:10.1186/1471-2180-13-222