Figure 1.

The C. burnetii T4BSS is expressed during growth in ACCM. (A) Total C. burnetii RNA was harvested from ACCM cultures at 96 and 168 hpi. Transcripts for dotH, icmT, icmV and icmW are detectable only when reverse transcriptase (RT) is added to the 96 and 168 hpi RT-PCR reactions. (C) Represents a chromosomal DNA PCR-positive control. (B) Total C. burnetii protein was harvested from the 96 and 168 hpi cultures. Equal volumes of the uninoculated and spent media (ACCM) from the respective cultures were retained and concentrated to equal volumes. The loading volumes of the 96 and 168 hpi cell samples were normalized using the membrane protein Com1. Predicted molecular weights (kDa) are indicated on the left. Immunoblot was used to detect the predicted cytoplasmic protein DotB, the predicted inner membrane protein DotN and the predicted outer membrane protein DotH. These proteins were detectable in all of the C. burnetii lysates and could not be detected in the ACCM.