title: Genetic transformation and phylogeny of bacterial symbionts from tsetse.
abstract: Two isolates of bacterial endosymbionts, GP01 and GM02, were established in cell free medium from haemolymph of the tsetse, Glossina pallidipes and G. morsitans. These microorganisms appear similar to rickettsia-like organisms reported previously from various tsetse species. The 16S rRNA sequence analysis, however, placed them within the gamma subdivision of the Proteobacteria, phylogenetically distinct from most members of the Rickettsiaceae which align with the alpha subdivision. Distinct multiple endogenous plasmids are harboured by GP01 and GM02, suggesting that the two isolates are different. Restriction mapping analysis showed that one of the conserved plasmids is present in high copy number and is at least 80 kb in size. A heterologous plasmid pSUP204, which contains the broad host range oriV replication origin, was used to transfect bacterial cultures. The symbiont GM02 was transformed, and it expressed plasmid encoded resistance to the antibiotics ampicillin, tetracycline and chloramphenicol. Transformation of these symbionts may provide a novel means for expressing anti-parasitic genes within tsetse populations.
authors.affiliation: Yale MacArthur Center for Molecular Parasitology, Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06510.
authors.forename: C B
authors.initials: CB
authors.lastname: Beard
authors.forename: S L
authors.initials: SL
authors.lastname: O'Neill
authors.forename: P
authors.initials: P
authors.lastname: Mason
authors.forename: L
authors.initials: L
authors.lastname: Mandelco
authors.forename: C R
authors.initials: CR
authors.lastname: Woese
authors.forename: R B
authors.initials: RB
authors.lastname: Tesh
authors.forename: F F
authors.initials: FF
authors.lastname: Richards
authors.forename: S
authors.initials: S
authors.lastname: Aksoy
mesh_terms: D000818:Animals; D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D006090:Gram-Negative Bacteria / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification / Q000648:ultrastructure; D008969:Molecular Sequence Data; D010957:Plasmids; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D013559:Symbiosis; D014170:Transformation, Genetic; D014345:Trypanosoma / Q000472:pathogenicity; D014352:Trypanosomiasis / Q000635:transmission; D014370:Tsetse Flies / Q000382:microbiology* / Q000469:parasitology / Q000648:ultrastructure
chemical_list: D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S