title: Transformation of Anaplasma phagocytophilum.
abstract: BACKGROUND
  Tick-borne pathogens cause emerging zoonoses, and include fastidious organisms such as Anaplasma phagocytophilum. Because of their obligate intracellular nature, methods for mutagenesis and transformation have not been available.
  
  
  RESULTS
  To facilitate genetic manipulation, we transformed A. phagocytophilum (Ap) to express a green fluorescent protein (GFP) with the Himar1 transposase system and selection with the clinically irrelevant antibiotic spectinomycin.
  
  
  CONCLUSIONS
  These transformed bacteria (GFP/Ap) grow at normal rates and are brightly fluorescent in human, monkey, and tick cell culture. Molecular characterization of the GFP/Ap genomic DNA confirmed transposition and the flanking genomic insertion locations were sequenced. Three mice inoculated with GFP/Ap by intraperitoneal injection became infected as demonstrated by the appearance of morulae in a peripheral blood neutrophil and re-isolation of the bacteria in culture.
authors.affiliation: Department of Entomology, University of Minnesota, St. Paul, MN 55108, USA. felsh001@umn.edu
authors.forename: Roderick F
authors.initials: RF
authors.lastname: Felsheim
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Herron
authors.forename: Curtis M
authors.initials: CM
authors.lastname: Nelson
authors.forename: Nicole Y
authors.initials: NY
authors.lastname: Burkhardt
authors.forename: Anthony F
authors.initials: AF
authors.lastname: Barbet
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Kurtti
authors.forename: Ulrike G
authors.initials: UG
authors.lastname: Munderloh
mesh_terms: D041081:Anaplasma phagocytophilum / Q000235:genetics* / Q000378:metabolism*; D000818:Animals; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D000882:Haplorhini; D006801:Humans; D013987:Ticks; D014169:Transformation, Bacterial / Q000502:physiology*
chemical_list: D049452:Green Fluorescent Proteins