In order to determine the localization and function of these C. burnetti proteins with the host cell, we constructed fusion proteins of each open reading frame(ORF) with mCherry, GFP, and FLAG peptides and expressed them in either eukaryotic cells or C. burnetti. We found the post infection CBU-1636 localized in the nucleus, CBU-1566 was more cytosolic and nuclear CBU-122 localized inside the CCV, and CBU-0885 was punctate nuclear and weak cytosolic. In order to prove that rach protein was a bona fide dot/icm effector, we constructed wildtype and icmL C. burnetti strains expressing effectors fused to andeylated cyclase. These strains will be assayed for cya-secretion into the cytosol. In conclusion, we observed noticeable differences in CBU-0122 and CBU-0885 post infection. In future studies, we hope to fix and stain the living image infected cells as a mean sof better comparison to the fixed and strained uninfected cells. We also hope to do the cya-assay to determine if these effector proteins are secreted by the dot/icm secretion system.
