Nonselective Persistence of a Rickettsia conorii Extrachromosomal Plasmid during Mammalian Infection
| Autor: | Abigail I. Fish, Fabio Del Piero, Sean P. Riley, Emma K. Harris, Daniel A. Garza, Juan J. Martinez, Kaikhushroo H. Banajee |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Male 030106 microbiology Immunology Virulence medicine.disease_cause Boutonneuse Fever Microbiology 03 medical and health sciences Mice Plasmid Transformation Genetic Bacterial Proteins Extrachromosomal DNA medicine Animals Humans Mice Inbred C3H biology Endothelial Cells Pathogenic bacteria Bacterial Infections biology.organism_classification Virology Spotted fever Transformation (genetics) Disease Models Animal Rickettsia conorii Infectious Diseases Rickettsia Parasitology Plasmids |
| Popis: | Scientific analysis of the genus Rickettsia is undergoing a rapid period of change with the emergence of viable genetic tools. The development of these tools for the mutagenesis of pathogenic bacteria will permit forward genetic analysis of Rickettsia pathogenesis. Despite these advances, uncertainty still remains regarding the use of plasmids to study these bacteria in in vivo mammalian models of infection, namely, the potential for virulence changes associated with the presence of extrachromosomal DNA and nonselective persistence of plasmids in mammalian models of infection. Here, we describe the transformation of Rickettsia conorii Malish 7 with the plasmid pRam18dRGA[AmTrCh]. Transformed R. conorii stably maintains this plasmid in infected cell cultures, expresses the encoded fluorescent proteins, and exhibits growth kinetics in cell culture similar to those of nontransformed R. conorii . Using a well-established murine model of fatal Mediterranean spotted fever, we demonstrate that R. conorii (pRam18dRGA[AmTrCh]) elicits the same fatal outcomes in animals as its untransformed counterpart and, importantly, maintains the plasmid throughout infection in the absence of selective antibiotic pressure. Interestingly, plasmid-transformed R. conorii was readily observed both in endothelial cells and within circulating leukocytes. Together, our data demonstrate that the presence of an extrachromosomal DNA element in a pathogenic rickettsial species does not affect either in vitro proliferation or in vivo infectivity in models of disease and that plasmids such as pRam18dRGA[AmTrCh] are valuable tools for the further genetic manipulation of pathogenic rickettsiae. |
| Databáze: | OpenAIRE |
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