title: Electrotransformation and Clonal Isolation of Rickettsia Species.
abstract: Genetic manipulation of obligate intracellular bacteria of the genus Rickettsia is currently undergoing a rapid period of change. The development of viable genetic tools, including replicative plasmids, transposons, homologous recombination, fluorescent protein-encoding genes, and antibiotic selectable markers has provided the impetus for future research development. This unit is designed to coalesce the basic methods pertaining to creation of genetically modified Rickettsia. The unit describes a series of methods, from inserting exogenous DNA into Rickettsia to the final isolation of genetically modified bacterial clones. Researchers working towards genetic manipulation of Rickettsia or similar obligate intracellular bacteria will find these protocols to be a valuable reference.
authors.affiliation: Vector-Borne Disease Laboratories, Department of Pathobiological Sciences, Louisiana State University School of Veterinary Medicine, Baton Rouge, Louisiana.
authors.forename: Sean P
authors.initials: SP
authors.lastname: Riley
authors.affiliation: Vector-Borne Disease Laboratories, Department of Pathobiological Sciences, Louisiana State University School of Veterinary Medicine, Baton Rouge, Louisiana.
authors.forename: Kevin R
authors.initials: KR
authors.lastname: Macaluso
authors.affiliation: Vector-Borne Disease Laboratories, Department of Pathobiological Sciences, Louisiana State University School of Veterinary Medicine, Baton Rouge, Louisiana.
authors.forename: Juan J
authors.initials: JJ
authors.lastname: Martinez
mesh_terms: D018274:Electroporation / Q000379:methods*; D005827:Genetics, Microbial / Q000379:methods*; D008967:Molecular Biology / Q000379:methods*; D012281:Rickettsia / Q000235:genetics* / Q000302:isolation & purification*; D014169:Transformation, Bacterial
keywords: cell sorting; clonal isolation; immunofluorescence; intracellular bacteria; limiting dilution; plaque assay; transformation