A method for purifying obligate intracellular Coxiella burnetii that employs digitonin lysis of host cells.
Purification of the obligate intracellular bacterium Coxiella burnetii requires physical disruption of infected cells. Here we describe a gentle and safe digitonin lysis procedure to release C. burnetii from infected cells. The purity, yield, and infectivity of digitonin-prepped organisms are comparable to that of organisms purified using cell lysis by sonication.
Cockrell
Diane C
DC
National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rocky Mountain Laboratories, Hamilton, Montana 59840, USA.
Beare
Paul A
PA
Fischer
Elizabeth R
ER
Howe
Dale
D
Heinzen
Robert A
RA
D000818:Animals; D001431:Bacteriological Techniques / Q000379:methods*; D002462:Cell Membrane / Q000187:drug effects; D002522:Chlorocebus aethiops; D016997:Coxiella burnetii / Q000166:cytology / Q000302:isolation & purification* / Q000648:ultrastructure; D004072:Digitonin / Q000378:metabolism*; D008853:Microscopy; D046529:Microscopy, Electron, Transmission; D014709:Vero Cells
D004072:Digitonin