Characterization And Cloning Of Coxiella burnetii

Abstract

Coxiella burnetii, an obligate intracellular parasite, is the etiological agent of the disease QFever, which can occur in acute and chronic forms. The acute form is rarely serious, but chronic Q-Fever may result in endocarditis, which is usually fatal. It is therefore of interest to determine which strains of Coxiella burnetii lead to the acute versus the endocarditis-associated sickness, and what the mechanisms involved in each are. This past summer at Rocky Mountain Laboratories in Hamilton, Montana, under the direction of Dr.Ted Hackstadt,I began to research the Coxiella burnetii 9-Mile strain, which is associated with the acute form of the disease. DNA characterization of this bacterium by restriction mapping was performed as well as attempts with antibodies to determine which proteins were cloned into recombinants. These could be compared to those of endocarditis-associated strains. With lambda gtll as a cloning and expression vector, DNA was inserted into various plasmids, proteins of the systems were expressed, and ligations and transformations of plasmids with Escherichia coli strains were carried out. These transformants were then used in plasmid mini-preparations for isolation and identification of the DNA we were successful in cloning.